title: Monogamy

text: Paleoanthropological estimates of the time frame for the evolution of monogamy are primarily based on the level of sexual dimorphism seen in the fossil record because, in general, the reduced male-male competition seen in monogamous mating results in reduced sexual dimorphism. According to Reno "et al.", the sexual dimorphism of "Australopithecus afarensis", a human ancestor from approximately 3.9–3.0 million years ago, was within the modern human range, based on dental and postcranial morphology. Although careful not to say that this indicates monogamous mating in early hominids, the authors do say that reduced levels of sexual dimorphism in "A. afarensis" "do not imply that monogamy is any less probable than polygyny". However, Gordon, Green and Richmond claim that in examining postcranial remains, "A. afarensis" is more sexually dimorphic than modern humans and chimpanzees with levels closer to those of orangutans and gorillas. Furthermore, "Homo habilis", living approximately 2.3 mya, is the most sexually dimorphic early hominid. Plavcan and van Schaik conclude their examination of this controversy by stating that, overall, sexual dimorphism in australopithecines is not indicative of any behavioral implications or mating systems.


title: Monogamy

text: Despite the human ability to avoid sexual and genetic monogamy, social monogamy still forms under many different conditions, but most of those conditions are consequences of cultural processes. These cultural processes may have nothing to do with relative reproductive success. For example, anthropologist Jack Goody's comparative study utilizing the "Ethnographic Atlas" demonstrated that monogamy is part of a cultural complex found in the broad swath of Eurasian societies from Japan to Ireland that practice social monogamy, sexual monogamy and dowry (i.e. "diverging devolution", that allow property to be inherited by children of both sexes). Goody demonstrates a statistical correlation between this cultural complex and the development of intensive plough agriculture in those areas. Drawing on the work of Ester Boserup, Goody notes that the sexual division of labour varies in intensive plough agriculture and extensive shifting horticulture. In plough agriculture farming is largely men's work and is associated with private property; marriage tends to be monogamous to keep the property within the nuclear family. Close family (endogamy) are the preferred marriage partners to keep property within the group. A molecular genetic study of global human genetic diversity argued that sexual polygyny was typical of human reproductive patterns until the shift to sedentary farming communities approximately 10,000 to 5,000 years ago in Europe and Asia, and more recently in Africa and the Americas. A further study drawing on the Ethnographic Atlas showed a statistical correlation between increasing size of the society, the belief in "high gods" to support human morality, and monogamy. A survey of other cross-cultural samples has confirmed that the absence of the plough was the only predictor of polygamy, although other factors such as high male mortality in warfare (in non-state societies) and pathogen stress (in state societies) had some impact.


title: Monogamy

text: Betzig postulated that culture/society can also be a source of social monogamy by enforcing it through rules and laws set by third-party actors, usually in order to protect the wealth or power of the elite. For example, Augustus Caesar encouraged marriage and reproduction to force the aristocracy to divide their wealth and power among multiple heirs, but the aristocrats kept their socially monogamous, legitimate children to a minimum to ensure their legacy while having many extra-pair copulations. Similarly—according to Betzig—the Christian Church enforced monogamy because wealth passed to the closest living, legitimate male relative, often resulting in the wealthy oldest brother being without a male heir. Thus, the wealth and power of the family would pass to the "celibate" younger brother of the church. In both of these instances, the rule-making elite used cultural processes to ensure greater reproductive fitness for themselves and their offspring, leading to a larger genetic influence in future generations. According to B. S. Low, culture would appear to have a much larger impact on monogamy in humans than the biological forces that are important for non-human animals.


title: Monogamy

text: Other theorists use cultural factors influencing reproductive success to explain monogamy. During times of major economic/demographic transitions, investing more in fewer offspring (social monogamy not polygyny) increases reproductive success by ensuring the offspring themselves have enough initial wealth to be successful. This is seen in both England and Sweden during the industrial revolution and is currently being seen in the modernization of rural Ethiopia. Similarly, in modern industrialized societies, fewer yet better-invested offspring, i.e. social monogamy, can provide a reproductive advantage over social polygyny, but this still allows for serial monogamy and extra-pair copulations.


title: Monogamy

text: Karol Wojtyła (later, Pope John Paul II) in his book "Love and Responsibility" postulated that monogamy, as an institutional union of two people being in love with one another, was an embodiment of an ethical "personalistic norm", and thus the only means of making true human love possible. Some writers have suggested that monogamy may solve the problems they view as associated with non-monogamy and hypergamy such as inceldom.


title: Monogamy

text: Alexandra Kollontai in "Make Way for the Winged Eros" argues that monogamy is an artifact of capitalist concepts of property and inheritance and wrote, "The social aims of the working class are not affected one bit by whether love takes the form of a long and official union or is expressed in a temporary relationship. The ideology of the working class does not place any formal limits on love." Later, "Modern love always sins, because it absorbs the thoughts and feelings of 'loving hearts' and isolates the loving pair from the collective. In the future society, such a separation will not only become superfluous but also psychologically inconceivable." One of the tenets of the new proletarian morality is "mutual recognition of the rights of the other, of the fact that one does not own the heart and soul of the other (the sense of property, encouraged by bourgeois culture)."


title: Monogamy

text: The historical record offers contradictory evidence on the development and extent of monogamy as a social practice. Laura Betzig argues that in the six large, highly stratified early states, commoners were generally monogamous but that elites practiced de facto polygyny. Those states included Mesopotamia, Egypt, Aztec Mexico, Inca Peru, India and China.


title: Monogamy

text: Monogamy has appeared in some traditional tribal societies such as the Andamanese, Karen in Burma, Sami and Ket in northern Eurasia, and the Pueblo Indians of the United States, apparently unrelated to the development of the Judeo-Christian monogamous paradigm.


title: Monogamy

text: Both the Babylonian and Assyrian families were monogamous in principle but not entirely so in practice since polygyny was frequently practiced by the rulers.


title: Monogamy

text: In the patriarchal society of Mesopotamia the nuclear family was called a "house". In order "to build a house" a man was supposed to marry one woman and if she did not provide him with offspring, he could take a second wife. The Code of Hammurabi states that he loses his right to do so if the wife herself gives him a slave as concubine. According to Old Assyrian texts, he could be obliged to wait for two or three years before he was allowed to take another wife. The position of the second wife was that of a "slave girl" in respect to the first wife, as many marriage contracts explicitly state.


title: Monogamy

text: Although an Egyptian man was free to marry several women at a time, and some wealthy men from Old and Middle Kingdoms did have more than one wife, monogamy was the norm. There may have been some exceptions, e.g. a Nineteenth Dynasty official stated as proof of his love to his deceased wife that he had stayed married to her since their youth, even after he had become very successful (P. Leiden I 371). This may suggest that some men abandoned first wives of a low social status and married women of higher status in order to further their careers although even then they lived with only one wife. Egyptian women had the right to ask for a divorce if their husband took a second wife. Many tomb reliefs testify to the monogamous character of Egyptian marriages; officials are usually accompanied by a supportive wife. "His wife X, his beloved" is the standard phrase identifying wives in tomb inscriptions. The instruction texts belonging to wisdom literature, e.g., Instruction of Ptahhotep or Instruction of Any, support fidelity to monogamous marriage life, calling the wife a "Lady of the house". The Instruction of Ankhsheshonq suggests that it is wrong to abandon a wife because she is not capable of pregnancy.


title: Monogamy

text: As against Betzig's contention that monogamy evolved as a result of Christian socio-economic influence in the West, monogamy appeared widespread in the ancient Middle East much earlier. In Israel's pre-Christian era, an essentially monogamous ethos underlay the Jewish creation story (Gn 2) and the last chapter of Proverbs. During the Second Temple period (530 BCE to 70 CE), apart from an economic situation which supported monogamy even more than in earlier period, the concept of "mutual fidelity" between husband and wife was a quite common reason for strictly monogamous marriages. Some marriage documents explicitly expressed a desire for the marriage to remain monogamous. Examples of these documents were found in Elephantine. They resemble those found in neighbouring Assyria and Babylonia. Study shows that ancient Middle East societies, though not strictly monogamous, were practically (at least on commoners' level) monogamous. Halakha of the Dead Sea Sect saw prohibition of polygamy as coming from the Pentateuch (Damascus Document 4:20–5:5, one of the Dead Sea Scrolls). Christianity adopted a similar attitude (cf. 1 Tm 3:2,12; Tt 1:6), which conformed with Jesus' approach. Michael Coogan, in contrast, states that "Polygyny continued to be practised well into the biblical period, and it is attested among Jews as late as the second century CE."


title: Monogamy

text: Under Judges and the monarchy, old restrictions went into disuse, especially among royalty, though the Books of Samuel and Kings, which cover entire period of monarchy, do not record a single case of bigamy among commoners—except for Samuel's father. The wisdom books e.g. Book of Wisdom, which provides a picture of the society, Sirach, Proverbs, Qohelet portray a woman in a strictly monogamous family (cf. Pr 5:15-19; Qo 9:9; Si 26:1-4 and eulogy of perfect wife, Proverbs 31:10-31). The Book of Tobias speaks solely of monogamous marriages. Also prophets have in front of their eyes monogamous marriage as an image of the relationship of God and Israel. (Cf. Ho 2:4f; Jer 2:2; Is 50:1; 54:6-7; 62:4-5; Ez 16). Roland de Vaux states that "it is clear that the most common form of marriage in Israel was monogamy".


title: Monogamy

text: The Mishnah and the "baraitot" clearly reflect a monogamist viewpoint within Judaism (Yevamot 2:10 etc.). Some sages condemned marriage to two wives even for the purpose of procreation (Ketubot 62b). R. Ammi, an "amora" states:


title: Monogamy

text: Whoever takes a second wife in addition to his first one shall divorce the first and pay her "kettubah" (Yevamot 65a)


title: Monogamy

text: Roman customs, which prohibited polygamy, may have enhanced such an attitude - especially after 212 AD, when all the Jews became Roman citizens. However, some Jews continued to practice bigamy (e.g. up to medieval times in Egypt and Europe). Fourth-century Roman law forbade Jews to contract plural marriages.


title: Monogamy

text: A synod convened by Gershom ben Judah around 1000 CE banned polygamy among Ashkenazi and Sephardic Jews.


title: Monogamy

text: The ancient Greeks and Romans were monogamous in the sense that men were not allowed to have more than one wife or to cohabit with concubines during marriage.


title: Monogamy

text: As John Paul II interpreted the dialogue between Jesus and the Pharisees (Gospel of Matthew 19:3–8), Christ emphasized the primordial beauty of monogamic spousal love described in the Book of Genesis 1:26–31, 2:4–25, whereby a man and woman by their nature are each ready to be a beautifying, total and personal gift to one another:


title: Monogamy

text: Jesus avoids entangling himself in juridical or casuistic controversies; instead, he appeals twice to the "beginning". By doing so, he clearly refers to the relevant words of Genesis, which his interlocutors also know by heart. ... it clearly leads the interlocutors to reflect about the way in which, in the mystery of creation, man was formed precisely as "male and female," in order to understand correctly the normative meaning of the words of Genesis.


title: Monogamy

text: Western European societies established monogamy as their marital norm. Monogamous marriage is normative and is legally enforced in most developed countries. Laws prohibiting polygyny were adopted in Japan (1880), China (1953), India (1955) and Nepal (1963). Polyandry is illegal in most countries.


title: Monogamy

text: The women's rights movements seek to make monogamy the only legal form of marriage. The United Nations General Assembly in 1979 adopted the Convention on the Elimination of All Forms of Discrimination Against Women, Article 16 of which requires nations to give women and men equal rights in marriage. Polygamy is viewed as inconsistent with the Article as it gives men the right of multiple wives, but not to women. The United Nations has established the Committee on the Elimination of Discrimination against Women (CEDAW) to monitor the progress of nations implementing the convention.


title: Monogamy

text: The founders of Communism determined that monogamous marriage inherently oppressed women and therefore had no place in communist society. Friedrich Engels stated that compulsory monogamy could only lead to increased prostitution and general immorality, with the benefits of restricting capital and solidifying the class structure. As he spelled out in "The Origin of the Family, Private Property and the State" (1884),


title: Monogamy

text: The first class antagonism which appears in history coincides with the development of the antagonism between man and woman in monogamian marriage, and the first class oppression with that of the female sex by the male. ... [T]he wellbeing and development of the one group are attained by the misery and repression of the other.


title: Monogamy

text: The monogamous family is distinguished from the pairing family by the far greater durability of wedlock, which can no longer be dissolved at the pleasure of either party. As a rule, it is only the man who can still dissolve it and cast off his wife.


title: Monogamy

text: However, the communist revolutionaries in China chose to take the Western viewpoint of monogamy as giving women and men equal rights in marriage. The newly formed Communist government established monogamy as the only legal form of marriage.


title: Monogamy

text: "The 1950 Marriage Law called for sweeping changes in many areas of family life. It forbade any 'arbitrary and compulsory' form of marriage that would be based on the superiority of men and would ignore women's interests. The new democratic marriage system was based on the free choice of couples, monogamy, equal rights for both sexes, and the protection of the lawful interests of women. It abolished the begetting of male offspring as the principal purpose of marriage and weakened kinship ties which reduced the pressure on women to bear many children, especially sons. With arranged marriages prohibited, young women could choose their own marriage partners, share the financial cost of setting up a new household, and have equal status in household and family decision-making. The Government then initiated an extensive campaign of marriage-law education, working jointly with the Communist Party, women's federations, trade unions, the armed forces, schools and other organizations."


title: Monogamy

text: The African Union has adopted the "Protocol on the Rights of Women in Africa" (the Maputo Protocol). While the protocol does not suggest making polygamous marriage illegal, Article 6 does state that "monogamy is encouraged as the preferred form of marriage and that the rights of women in marriage and family, including in polygamous marital relationships are promoted and protected." The protocol entered into force on 25 November 2005.


title: Monogamy

text: Recent discoveries have led biologists to talk about the three varieties of monogamy: social monogamy, sexual monogamy, and genetic monogamy. The distinction between these three are important to the modern understanding of monogamy.


title: Monogamy

text: Monogamous pairs of animals are not always sexually exclusive. Many animals that form pairs to mate and raise offspring regularly engage in sexual activities with partners other than their primary mate. This is called extra-pair copulation. Sometimes these extra-pair sexual activities lead to offspring. Genetic tests frequently show that some of the offspring raised by a monogamous pair come from the female mating with an extra-pair male partner. These discoveries have led biologists to adopt new ways of talking about monogamy:


title: Monogamy

text: Whatever makes a pair of animals socially monogamous does not necessarily make them sexually or genetically monogamous. Social monogamy, sexual monogamy, and genetic monogamy can occur in different combinations.


title: Monogamy

text: Social monogamy does not always involve marriage in humans. A married couple is almost always a socially monogamous couple. But couples who choose to cohabit without getting married can also be socially monogamous. The popular science author Matt Ridley in his book "The Red Queen: Sex and the Evolution of Human Nature", described the human mating system as "monogamy plagued by adultery".


title: Monogamy

text: Serial monogamy is a mating practice in which individuals may engage in sequential monogamous pairings, or in terms of humans, when men or women can marry another partner but only after ceasing to be married to the previous partner.


title: Monogamy

text: Serial monogamy may effectively resemble polygyny in its reproductive consequences because some men are able to utilize more than one woman's reproductive lifespan through repeated marriages.


title: Monogamy

text: Serial monogamy may also refer to sequential sexual relationships, irrespective of marital status. A pair of humans may remain sexually exclusive, or monogamous, until the relationship has ended and then each may go on to form a new exclusive pairing with a different partner. This pattern of serial monogamy is common among people in Western cultures.


title: Monogamy

text: Evolutionary theory predicts that males would be apt to seek more mating partners than females because they obtain higher reproductive benefits from such a strategy. Men with more serial marriages are likely to have more children than men with only one spouse, whereas the same is not true of women with consecutive spouses. A study done in 1994 found that remarried men often had a larger age difference from their spouses than men who were married for the first time, suggesting that serial monogamy helps some men extract a longer reproductive window from their spouses.


title: Monogamy

text: Serial monogamy has always been closely linked to divorce practices. Whenever procedures for obtaining divorce have been simple and easy, serial monogamy has been found. As divorce has continued to become more accessible, more individuals have availed themselves of it, and many go on to remarry. Barry Schwartz, author of "The Paradox of Choice: Why More is Less", further suggests that Western culture's inundation of choice has devalued relationships based on lifetime commitments and singularity of choice. It has been suggested, however, that high mortality rates in centuries past accomplished much the same result as divorce, enabling remarriage (of one spouse) and thus serial monogamy.


title: Monogamy

text: Monogamy is one of several mating systems observed in animals. However, a pair of animals may be socially monogamous without necessarily being sexually or genetically monogamous. Social monogamy, sexual monogamy, and genetic monogamy can occur in different combinations.


title: Monogamy

text: Social monogamy refers to the overtly observed living arrangement whereby a male and female share territory and engage in behaviour indicative of a social pair, but does not imply any particular sexual fidelity or reproductive pattern. The extent to which social monogamy is observed in animals varies across taxa, with over 90 percent of avian species being socially monogamous, compared to only 3 percent of mammalian species and up to 15 percent of primate species. Social monogamy has also been observed in reptiles, fish, and insects.


title: Monogamy

text: Sexual monogamy is defined as an exclusive sexual relationship between a female and a male based on observations of sexual interactions. However, scientific analyses can test for paternity, for example by DNA paternity testing or by fluorescent pigment powder tracing of females to track physical contact. This type of analysis can uncover reproductively successful sexual pairings or physical contact. Genetic monogamy refers to DNA analyses confirming that a female-male pair reproduce exclusively with each other.


title: Monogamy

text: The incidence of sexual monogamy appears quite rare in other parts of the animal kingdom. It is becoming clear that even animals that are overtly socially monogamous engage in extra-pair copulations. For example, while over 90% of birds are socially monogamous, "on average, 30% or more of the baby birds in any nest [are] sired by someone other than the resident male." Patricia Adair Gowaty has estimated that, out of 180 different species of socially monogamous songbirds, only 10% are sexually monogamous. Offspring are far more successful when both the male and the female members of the social pair contribute food resources.


title: Monogamy

text: The highest known frequency of reproductively successful extra-pair copulations are found among fairywrens "Malurus splendens" and "Malurus cyaneus" where more than 65% of chicks are fathered by males outside the supposed breeding pair. This discordantly low level of genetic monogamy has been a surprise to biologists and zoologists, as social monogamy can no longer be assumed to determine how genes are distributed in a species.


title: Monogamy

text: "Elacatinus", also widely known as neon gobies, also exhibit social monogamy. Hetereosexual pairs of fish belonging to the genus "Elacatinus" remain closely associated during both reproductive and non-reproductive periods, and often reside in same cleaning station to serve client fish. Fish of this genus frequently mate with a new partner after they are widowed.


title: Monogamy

text: Socially monogamous species are scattered throughout the animal kingdom: A few insects, a few fish, about nine-tenths of birds, and a few mammals are socially monogamous. There is even a parasitic worm, "Schistosoma mansoni", that in its female-male pairings in the human body is monogamous. The diversity of species with social monogamy suggests that it is not inherited from a common ancestor but instead evolved independently in many different species.


title: Monogamy

text: The low occurrence of social monogamy in placental mammals has been claimed to be related to the presence or absence of estrus—or oestrus—the duration of sexual receptivity of a female. This, however, does not explain "why" estrus females generally mate with any proximate male nor any correlation between sexual and social monogamy. Birds, which are notable for a high incidence of social monogamy, do not have estrus.


title: Monogamy

text: The prairie vole is an animal example for its monogamous social behaviour, since the male is usually socially faithful to the female, and shares in the raising of pups. The woodland vole is also usually monogamous. Another species from the same genus, the meadow vole, has promiscuously mating males, and scientists have changed adult male meadow voles' behaviour to resemble that of prairie voles in experiments in which a single gene was introduced into the brain by a virus.


title: Monogamy

text: The behaviour is influenced by the number of repetitions of a particular string of microsatellite DNA. Male prairie voles with the longest DNA strings spend more time with their mates and pups than male prairie voles with shorter strings. However, other scientists have disputed the gene's relationship to monogamy, and cast doubt on whether the human version plays an analogous role. Physiologically, pair-bonding behavior has been shown to be connected to vasopressin, dopamine, and oxytocin levels, with the genetic influence apparently arising via the number of receptors for these substances in the brain; the pair-bonding behavior has also been shown in experiments to be strongly modifiable by administering some of these substances directly.


title: Monogamy

text: The North American microtine rodent's (vole) complex social structure and social behavior has provided unique opportunities to study the underlying neural bases for monogamy and social attachment. Data from studies using the "Microtus ochrogaster" or prairie vole indicate that the neuroendocrine hormones, oxytocin (in female prairie voles) and vasopressin (in male prairie voles) play a central role in the development of affiliative connections during mating. The effects of intracerebroventricular administration of oxytocin and vasopressin have been shown to promote affiliative behavior in the prairie vole but not in similar, but non-monogamous montane voles. This difference in neuropeptide effect is attributed to the location, density, and distribution of OT and AVP receptors. Only in the prairie voles are OT and AVP receptors located along the mesolimbic dopamine reward pathway, presumably conditioning the voles to their mates odor while consolidating the social memory of the mating episode. This finding highlights the role of genetic evolution in altering the neuroanatomical distribution of receptors, resulting in certain neural circuits becoming sensitive to changes in neuropeptides.


title: Evelyn Nesbit

text: Evelyn Nesbit (born Florence Evelyn Nesbit; December 25, 1884 or 1885 – January 17, 1967) was an American artists' model, chorus girl, and actress. She is best known for her years as a young woman in New York City, particularly her involvement in a deadly and abusive triangle between railroad scion Harry Kendall Thaw and architect Stanford White, which resulted in White's murder by Thaw in 1906.


title: Evelyn Nesbit

text: In her day, Nesbit was a famous fashion model, being frequently photographed for mass circulation newspapers, magazine advertisements, souvenir items, and calendars. When in her early teens, she had begun working as an artist's model in Philadelphia. Nesbit continued after her family moved to New York, posing for legitimate artists including James Carroll Beckwith, Frederick S. Church, and notably Charles Dana Gibson, who idealized her as a "Gibson Girl". She was an artists' and fashion model when both fashion photography (as an advertising medium) and the pin-up (as an art genre) were beginning to expand.


title: Evelyn Nesbit

text: Nesbit entered theater, first as a chorus line dancer. She was offered a contract as a featured star, and attracted attention from a variety of wealthy men, including Stanford White, nearly three times her age. In 1905, Nesbit married Harry Thaw, a multi-millionaire with a history of mental instability and abusive behavior. The next year, on June 25, 1906, Thaw shot and killed White at the rooftop theatre of Madison Square Garden.


title: Evelyn Nesbit

text: The press called the resulting court case the "Trial of the Century", and coverage of these well-known figures was sensational. Nesbit testified that White had befriended her and her mother, but had allegedly drugged her and then raped her when she was unconscious. Nesbit and White had also had an ongoing relationship after the alleged rape incident. Thaw was said to have killed White in retaliation for his actions with Nesbit, based on his own obsession with her.


title: Evelyn Nesbit

text: Nesbit visited Thaw while he was confined to mental asylums. She toured Europe with a dance troupe, and her son, Russell Thaw, was born there. Later she took the boy with her to Hollywood, where she appeared as an actress in numerous silent films. Russell also acted in some of these. Nesbit wrote two memoirs about her life, published in 1914 and 1934. Her son later became a pilot, working as a test pilot for the Douglas Aircraft Company after World War II.


title: Evelyn Nesbit

text: Florence Evelyn Nesbit was born on December 25, 1884, or December 25, 1885, in Natrona, Pennsylvania, a small town near Pittsburgh. In her childhood, she was primarily known as Florence Evelyn. The year of her birth remains unconfirmed, as the local records were later destroyed in a fire and Evelyn said she was unsure of it; some sources have put the year as 1884, some as 1885, and it could have been something else. In later years, Nesbit confirmed that her mother at times added several years to her age as a girl to circumvent child labor laws.


title: Evelyn Nesbit

text: Nesbit was the daughter of Winfield Scott Nesbit and his wife, Evelyn Florence (née McKenzie), and was of Scots-Irish ancestry. Her father was an attorney and her mother was a homemaker. Nesbit later said that she had an especially close relationship with her father, and tried to please him by her accomplishments. Mr. Nesbit encouraged her curiosity and self-confidence. As she loved reading, he chose books for her and set up a small library for her use, consisting of fairy tales, fantasies, and also books regarded typically as of interest to boys only – the "pluck and luck" stories that were popular in that era. When Nesbit showed an interest in music and dance, her father encouraged her to take lessons.


title: Evelyn Nesbit

text: The Nesbit family moved to Pittsburgh around 1893. When Nesbit was about 10, her father died suddenly at age 40, leaving the family penniless. Her family lost their home, and all their possessions were auctioned off to pay outstanding debts. Nesbit's mother was unable to find work using her dressmaking skills, and the family had to depend on the charity of friends and relatives. The family lived as nomads, sharing a single room in a series of boarding houses. Her mother often sent Nesbit's younger brother Howard to live with relatives or family friends for periods of time. Nesbit's mother was eventually given money to rent a house to use as a boardinghouse, in order to have a source of income. She sometimes assigned young Evelyn (aged about 12) to the duty of collecting the rent from boarders. In her 1915 memoir, Nesbit later recalled that, "Mamma was always worried about the rent  ... it was too hard a thing for her to actually ask for every week, and it never went smoothly." Nesbit's mother lacked the temperament or savvy to run a boardinghouse, and the venture failed.


title: Evelyn Nesbit

text: Under continuous financial distress which showed no prospect of improvement, Mrs. Nesbit moved to Philadelphia in 1898. A friend had encouraged her, advising that relocation to Philadelphia could open opportunities for her employment as a seamstress. Evelyn and Howard were sent to an aunt and then transferred for care to a family in Allegany, whose acquaintance their mother had made some years earlier. Mrs. Nesbit indeed gained a job, not as a seamstress, but as a sales clerk at the fabric counter of Wanamaker's department store. She sent for her children, and both 14-year-old Evelyn and 12-year-old Howard also became Wanamaker employees, working twelve-hour days for six days a week.


title: Evelyn Nesbit

text: It was here that Nesbit had a chance encounter with an artist who was struck by her beauty. She asked Nesbit to pose for a portrait, which her mother agreed to after verifying the artist was a woman. Nesbit sat for five hours and earned one dollar (equivalent to approximately $27.50 in 2016). She was introduced to other artists in the Philadelphia area, and became a favorite model for a group of reputable illustrators, portrait painters, and stained-glass artisans. In later life, Nesbit explained: "When I saw I could earn more money posing as an artist's model than I could at Wanamaker's, I gave my mother no peace until she permitted me to pose for a livelihood."


title: Evelyn Nesbit

text: In June 1900, Mrs. Nesbit, leaving her children in the care of others, relocated to New York City to seek work as a seamstress or clothing designer. However, she did not succeed in this competitive world. In November 1900, she finally sent for her children, although she had no work. The family shared a single back room in a building on 22nd Street in Manhattan.


title: Evelyn Nesbit

text: Mrs. Nesbit finally used letters of introduction given by Philadelphia artists, contacting painter James Carroll Beckwith. His primary patron was John Jacob Astor. Beckwith was both a respected painter and instructor of life classes at the Art Students League. He took a protective interest in the young Nesbit, and provided her with letters of introduction to other legitimate artists, such as Frederick S. Church, Herbert Morgan, and Carle J. Blenner.


title: Evelyn Nesbit

text: Mrs. Nesbit was forced to take on managing her daughter's career, proving unable to provide either business acumen or guardianship for her daughter. In a later interview with reporters, Mrs. Nesbit maintained: "I never allowed Evelyn to pose in the altogether". Two artworks, one by Church and another by Beckwith in 1901, contradict her statement, as they display a skimpily clad or partially nude Evelyn.


title: Evelyn Nesbit

text: Nesbit became one of the most in-demand artists' models in New York. Photographers Otto Sarony and Rudolf Eickemeyer were among those who worked with her. She was featured on the covers of numerous women's magazines of the period, including "Vanity Fair", "Harper's Bazaar", "The Delineator", "Ladies' Home Journal", and "Cosmopolitan". She also appeared in fashion advertising for a wide variety of products; and she was also showcased on sheet music and souvenir items – beer trays, tobacco cards, pocket mirrors, postcards, and chromolithographs. Nesbit often posed in vignettes, dressed in various costumes. These photo postcards were known as "mignon" (sweet, lovely), as their pictorials were of a suggestive sensuality in contrast to the graphic, notorious "French postcards" of the day. She also posed for calendars for Prudential Life Insurance, Coca-Cola, and other corporations. Charles Dana Gibson, one of the country's most renowned artists of the era, used Nesbit as the model for one of his best-known "Gibson Girl" works. Titled "Woman: The Eternal Question" (1903), the portrait features her in profile, with her luxuriant hair forming the shape of a question mark.


title: Evelyn Nesbit

text: The use of photographs of young women in advertising, referred to as the "live model" style, was just beginning to be widely used and began to supplant illustration. Nesbit modeled for Joel Feder, an early pioneer in fashion photography. She found such assignments less strenuous than working as an artist's model, as posing sessions were shorter. The work was lucrative. With Feder, Nesbit earned $5 for a half-day shoot and $10 for a full day – $ per day in dollars. Eventually, the fees she earned from her modeling career exceeded the combined income which her family had earned at Wanamaker's department store. But the high cost of living in New York strained their finances.


title: Evelyn Nesbit

text: Nesbit became disaffected and bored with the long hours spent in confined environments, maintaining the immobile poses required of a studio model. Her popularity in modeling had attracted the interests of theatrical promoters, some legitimate and some disreputable, who offered her acting opportunities. Nesbit pressed her mother to let her enter the theatre world, and Mrs. Nesbit ultimately agreed to let her daughter try this new way to augment their finances. An interview was arranged for the aspiring performer with John C. Fisher, company manager of the wildly popular play "Florodora", then enjoying a long run at the Casino Theatre on Broadway. Mrs. Nesbit's initial objections were softened by the knowledge that some of the girls in the show had managed to marry millionaires. In July 1901, costumed as a "Spanish maiden", Nesbit became a member of the show's chorus line, whose enthusiastic public dubbed them the "Florodora Girls". Billed as "Florence Evelyn", the new chorus girl was called "Flossie the Fuss" by the cast, a nickname which displeased her. She changed her theatrical name to Evelyn Nesbit.


title: Evelyn Nesbit

text: After her stint with "Florodora" ended, Nesbit sought out other roles. She won a part in "The Wild Rose", which had just come to Broadway. After an initial interview with Nesbit, the show's producer, George Lederer, sensed he had discovered a new sensation. He offered her a contract for a year and, more significantly, moved her out of the chorus line and into a position as a featured player – the role of the Gypsy girl "Vashti". The publicity machine began to roll, possibly fueled by the influence of architect Stanford White, and she was hyped in the gossip columns and theatrical periodicals of the day. On May 4, 1902, "The New York Herald" showcased Nesbit in a two-page article, enhanced by photographs, promoting her rise as a new theatrical light, and recounting her career from model to chorus line to key cast member. "Her Winsome Face to be Seen Only from 8 to 11pm", the newspaper title announced to the public. The press coverage invariably touted her physical charms and potent stage presence; acting skills were rarely mentioned.


title: Evelyn Nesbit

text: As a chorus girl on Broadway in 1901, at the age of 15 or 16, Nesbit was introduced to White by Edna Goodrich, who was also a member of the company of "Florodora." White, known as "Stanny" by close friends and relatives, was 46 years old. Despite being married with a son, White had an independent social life. He used intermediaries to disarm the girl. Nesbit was initially struck by White's imposing size, which she said "was appalling", and she also said that to her he seemed "terribly old".


title: Evelyn Nesbit

text: White invited Nesbit and Goodrich to lunch at his multi-floor apartment on West 24th Street above the toy store FAO Schwarz, the entrance of which was next to the store's back delivery entrance. In her memoir "Prodigal Days" (1934), Nesbit described being overwhelmed by White's expensive furnishings and luxurious apartment. He had another male guest about his age, Reginald Ronalds. The luncheon was as extravagant as the setting. Afterward, the party went two flights up to a room decorated in green, where a large, red velvet swing was suspended from the ceiling. Nesbit agreed to sit in it, and White pushed her. The four played spontaneous games involving the swing.


title: Evelyn Nesbit

text: White appeared to be a witty, kind, and generous man. The wealthy socialite was described in newspapers as "masterful", "intense", and "burly yet boyish". He impressed both Nesbit and her mother as an "interesting companion". White sponsored Nesbit, her mother and brother for better living quarters, moving them into a suite at the Wellington Hotel, which he also furnished. White soon won over Mrs. Nesbit; in addition to providing the apartment, he paid for her son, Howard, to attend the Chester Military Academy near Philadelphia. He also persuaded Mrs. Nesbit to take a trip to visit friends in Pittsburgh, assuring her he would watch over her daughter Evelyn.


title: Evelyn Nesbit

text: While her mother was out of town, Nesbit and White had dinner and champagne at his apartment, capped by a tour ending at the "mirror room", which was furnished only with a green velvet sofa. After they drank more champagne, Nesbit changed into a yellow satin kimono at White's request. She later said this was her last memory. When Nesbit awoke, she found herself naked in bed next to an also-naked White and saw blood on the sheets, marking the loss of her virginity. Despite the apparent date rape, Nesbit allowed White to be her regular lover and close companion for some time. As their relationship faded, she discovered that he had had affairs with other young women whose names he recorded in a "little black book".


title: Evelyn Nesbit

text: John Barrymore became entranced with Nesbit's performance in "The Wild Rose" and attended the show at least a dozen times. The two met at a lavish party given by White, who had invited Barrymore, the brother of his friend, stage actress Ethel Barrymore. In 1902, a romance blossomed between Nesbit and Barrymore, a 21-year-old contemporary who was close to her own age. He was witty and fun-loving, and Nesbit became smitten with him. After an evening out, the couple often returned to Barrymore's apartment, staying until the early-morning hours. Barrymore was casually pursuing a career as illustrator and cartoonist. Although he showed some promise in his chosen field, his salary was small and he behaved irresponsibly with the family money. Nesbit's mother and White considered him an unsuitable match for Nesbit, and both were greatly displeased when they found out about the relationship.


title: Evelyn Nesbit

text: White worked to separate the couple by arranging for Nesbit's enrollment at a boarding school in New Jersey, administered by Mathilda DeMille, mother of film director Cecil B. DeMille. In the presence of both Mrs. Nesbit and White, Barrymore had asked Nesbit to marry him, but she turned him down.


title: Evelyn Nesbit

text: In 1939, Barrymore and Nesbit had a tearful reunion in Chicago. He was in town starring in “My Dear Children” and, one night after the show, found his way to Gene Harris’ Club Alabam, where she was appearing on stage. According to legend, Barrymore announced to the room that she was his first love.


title: Evelyn Nesbit

text: Aside from her relationship with Barrymore, Nesbit was involved with other men who vied for her attention. Among those were the polo player James Montgomery "Monte" Waterbury and the young magazine publisher Robert J. Collier. Even as she had these relationships, White still remained a potent presence in Nesbit's life and served as her benefactor.


title: Evelyn Nesbit

text: Nesbit eventually became involved with Harry Kendall Thaw, the son of a Pittsburgh coal and railroad baron. With a history of pronounced mental instability dating to his childhood, Thaw, heir to a $40 million fortune, led a reckless, self-indulgent life. He had attended some 40 performances of "The Wild Rose", over nearly a year. Even before he met Nesbit, Thaw had developed a resentment of White, believing that he had blocked Thaw's acceptance in social circles and was a womanizer who preyed on young women. Thaw may have chosen Nesbit because of her relationship with White.


title: Evelyn Nesbit

text: Through an intermediary, Thaw arranged a meeting with Nesbit, introducing himself as "Mr. Munroe". He maintained this subterfuge while giving her items and money. One day he confronted Evelyn and said: "I am not Munroe ... I am Harry Kendall Thaw, of Pittsburgh!" She did not react with such surprise as he had expected; she was already used to attracting the attention of wealthy men.


title: Evelyn Nesbit

text: In early 1903, while at boarding school, Nesbit underwent emergency surgery. The official diagnosis was acute appendicitis; however, some sources have speculated that she had been pregnant (perhaps by Barrymore) and had an abortion. Evelyn's grandson, Russell Thaw, has said: "I think she went away to have an abortion." However, under oath at Thaw's murder trials, both Barrymore and she denied that she was pregnant or had an abortion.


title: Evelyn Nesbit

text: Thaw became solicitous, ensuring that Nesbit received the best medical care available. He suggested that she should go on a European trip, convincing Nesbit and her mother that this would hasten the young woman's recovery. Evelyn's mother accompanied them for propriety. Thaw created a hectic itinerary and rate of travel. Tensions mounted between mother and daughter, and Mrs. Nesbit insisted on returning to the United States. Thaw took Nesbit alone to Paris, leaving Mrs. Nesbit in London. In Paris, he pressed Evelyn to become his wife, but she refused. Aware of Thaw's obsession with female chastity, she could not accept his marriage proposal without revealing the truth of her relationship with White. Thaw continued to interrogate her, and ultimately Nesbit told him of White's assault. Thaw accused her mother of being an unfit parent.


title: Evelyn Nesbit

text: Thaw and Evelyn continued their travel through Europe where he took her to sites devoted to the cult of virgin martyrdom. In Domrémy, France, the birthplace of Joan of Arc, Thaw wrote in the visitor's book: "she would not have been a virgin if Stanford White had been around." In Austria-Hungary, Thaw took Evelyn to the gothic , where he had the three servants in residence butler, cook, and maid kept at one end of the building; while he and Nesbit had isolated quarters at the opposite end. Thaw locked Evelyn in her room, then beat her with a whip and sexually assaulted her over a two-week period. Afterward, he was apologetic and upbeat.


title: Evelyn Nesbit

text: After returning to New York, Nesbit talked to friends about her ordeal. Others shared stories about Thaw and a propensity toward myriad addictive behaviors. Several men told her that Thaw "took morphine" and "he was crazy".


title: Evelyn Nesbit

text: Although White was still part of her life, Nesbit realized they had no future together. She also knew her relationship with him had already compromised her reputation; if the full extent of their involvement became common knowledge, no respectable man would make her his wife. Nesbit also resented White for failing to tell her about Thaw's excesses and derangement. As a teenager, Nesbit had spent her formative years thrust into the adult society of artists and theatre people; her development had proceeded without the companionship of contemporaries of her own age. Her mother had remarried, and although she had been an inept guardian before, their estrangement was now complete. Nesbit was desperate to escape the poverty which she and her family had long suffered.


title: Evelyn Nesbit

text: Thaw continued to pursue Nesbit for marriage, promising that following their union he would live the life of a "Benedictine monk". With a perverted sense of justice, and a show of magnanimous charity, he assured Nesbit he had forgiven her for her relationship with White. Nesbit finally consented to marry Thaw. His mother agreed to the marriage, on the condition that Nesbit give up the theatre and modeling, and refrain from talking about her past life.


title: Evelyn Nesbit

text: Nesbit married Thaw on April 4, 1905, in Pittsburgh. For her wedding dress, Thaw chose a black traveling suit decorated with brown trim. Newspapers announced that the new Mrs. Thaw was now the "Mistress of Millions". The two took up residence in the Thaw family home, Lyndhurst, in Pittsburgh. Isolated with Thaw's mother and her like-minded social group of strict Presbyterians, Nesbit became the proverbial bird in a gilded cage. In later years, she said that the Thaws had a shallow value system: "the plane of materialism which finds joy in the little things that do not matter – the of ... [things]".


title: Evelyn Nesbit

text: Nesbit had imagined travel and entertaining, but found that her husband acted as a pious son. Thaw started a campaign to expose White, corresponding with reformer Anthony Comstock, a crusader for moral probity and the expulsion of vice. He became convinced that he was being stalked by members of the notorious Monk Eastman Gang of New York, believing White had hired them. Nesbit later said: "[Thaw] imagined his life was in danger because of the work he was doing in connection with the vigilance societies and the exposures he had made to those societies of the happenings in White's flat." In reality, White, not thought to have been aware of Thaw's animus, considered him a poseur of little consequence, categorizing him as a clown and calling him the "Pennsylvania pug", a reference to Thaw's baby-faced features.


title: Evelyn Nesbit

text: Thaw and Nesbit visited New York in June 1906 before boarding a luxury liner bound for a European holiday. Late that day, Thaw said that he had tickets for them for the premiere of "Mam'zelle Champagne", written by Edgar Allan Woolf, at the rooftop theatre of Madison Square Garden. They first stopped at the Cafe Martin for dinner, where they happened to see White, before going to the theatre. Despite the heat, Thaw wore a long black overcoat over his tuxedo and refused to remove it.


title: Evelyn Nesbit

text: At 11:00 pm, as the stage show was coming to a close, White appeared and took his place at a table customarily reserved for him. Spotting his arrival, Thaw approached him several times, each time withdrawing. During the finale, "I Could Love A Million Girls", Thaw produced a pistol, and, from two feet away, fired three shots into White's head and back, killing him instantly. Thaw addressed the crowd, but witness reports varied as to his words. He said (roughly): "I did it because he ruined my wife! He had it coming to him! He took advantage of the girl and then abandoned her! ... You'll never go out with that woman again!" In his book "The Murder of Stanford White" (2011), Gerald Langford quoted Thaw as saying, "You ruined my life", or, "You ruined my wife."


title: Evelyn Nesbit

text: The crowd initially thought the incident might be a practical joke, but became alarmed upon realizing White was dead. Thaw brandished the pistol and was taken into police custody. Nesbit managed to extricate herself from the ensuing chaos on the Madison Square rooftop. Not wanting to return to their hotel suite, she took refuge for several days in the apartment of a chorus girl friend. Years later, Nesbit said of this time: "A complete numbness of mind and body took possession of me ... I moved like a person in a trance for hours afterward."


title: Evelyn Nesbit

text: As early as the morning following the murder, news coverage became both chaotic and single-minded, and ground forward with unrelenting momentum. A person, a place, or event, no matter how peripheral to the killing of White, was seized on by reporters and hyped as newsworthy copy. Facts were thin, but sensationalist reportage was plentiful in the heyday of yellow journalism. One week after the killing, the film "Rooftop Murder" was released for public viewing at the nickelodeon theaters, rushed into production by Thomas Edison.


title: Evelyn Nesbit

text: The hard-boiled male reporters of the yellow press were bolstered by a contingent of female counterparts, christened "Sob Sisters" or "The Pity Patrol." Initially, female spectators were allowed in to witness the proceedings. When the case came to trial, the judge banned women from the courtroom – excepting Thaw family members and the female news reporters there on "legitimate business".


title: Evelyn Nesbit

text: Female reporters wrote human interest pieces, emphasizing sentiment and melodrama. They were less sympathetic to Nesbit than Thaw. Nixola Greeley-Smith wrote of Nesbit: "I think that she was sold to one man [White] and later sold herself to another [Thaw]." In an article titled "The Vivisection of a Woman's Soul", Greeley-Smith described Nesbit's unmaidenly revelations as she testified on the stand: "Before her audience of many hundred men, young Mrs. Thaw was compelled to reveal in all its hideousness every detail of her association with Stanford White after his crime against her."


title: Evelyn Nesbit

text: The rampant interest in the White killing and its key players were used by both the defense and prosecution to feed malleable reporters any "scoops" that would give their respective sides an advantage in the public forum. News coverage dissected all the key players in what was called the "Garden Murder". One florid account keynoted Nesbit's vulnerability: "Her baby beauty proved her undoing. She toddled as innocently into the arms of Satan as an infant into the outstretched arms of parental love ..." Neither was her mother spared the scrutiny of rogue reporting: "She [her mother] knew better. She also knew she was sacrificing her child's soul for money ..."


title: Evelyn Nesbit

text: Church groups lobbied to restrict the media coverage, asking the government to step in as censor. President Theodore Roosevelt decried the newspapers' penchant for printing the "full disgusting particulars" of the trial proceedings. He conferred with the U.S. Postmaster General on the viability of prohibiting the dissemination of such printed matter through the United States mail, and censorship was threatened but never carried out.


title: Evelyn Nesbit

text: White was hounded in death, excoriated as a man and questioned as an architect. "The Evening Standard" concluded he was "more of an artist than architect"; his work spoke of his "social dissolution". "The Nation" was also critical: "He adorned many an American mansion with irrelevant plunder." Richard Harding Davis, a war correspondent and reputedly the model for the "Gibson Man", was angered by the yellow press, saying they had distorted the facts about his friend. "Vanity Fair" published an editorial lambasting White, which prompted Davis to write a rebuttal published in "Collier's", in which he attested that White "admired a beautiful woman as he admired every other beautiful thing God has given us; and his delight over one was as keen, as boyish, as grateful over any others."


title: Evelyn Nesbit

text: Thaw's mother was adamant that her son not be stigmatized by clinical insanity. She pressed for the defense to follow a compromise strategy: one of temporary insanity, or what in that era was referred to as a "brainstorm". Acutely conscious that her own family had a history of hereditary insanity, and after years of protecting her son's hidden life, she feared his past would be dragged out into the open, ripe for public scrutiny. Mrs. Thaw proceeded to hire a team of doctors, at a cost of some $500,000, to substantiate that her son's act of homicide constituted a single aberrant act. Nesbit in later years described the determination with which Thaw's family worked to favorably spin his mental deficiency: "the Thaws will put the biggest lunacy experts that money can buy on the stand ... Harry was a madman but they will prove it nicely".


title: Evelyn Nesbit

text: Again maneuvering her way through the gantlet of reporters, the curious public, the sketch artists, and photographers enlisted to capture the effect the "harrowing circumstances [had] on her beauty", Nesbit returned to her hotel and the assembled Thaw family. The Thaws may have promised Nesbit a comfortable financial future if she provided testimony at trial favorable to Thaw's case. It was a conditional agreement; if the outcome proved negative, she would receive nothing. The rumored amount of money the Thaws pledged for her cooperation ranged from $25,000 to $1,000,000.


title: Evelyn Nesbit

text: Nesbit's mother remained conspicuously absent throughout her daughter's entire ordeal. Mrs. Nesbit had been cooperating with the prosecution as Thaw's lawyers considered her culpable of prostituting her daughter to White. Nesbit's brother Howard, who had come to regard White as a father figure, blamed her for his death.


title: Evelyn Nesbit

text: Thaw was tried twice for the killing of White. Nesbit testified at both trials; her appearance on the witness stand was an emotionally tortuous ordeal. In open court, she was forced to detail her relationship with White, including the night when he allegedly raped her. Until then, the sexual assault had been a secret she had guarded at White's request. Other than White, only she and Thaw knew what had happened.


title: Evelyn Nesbit

text: Due to the unusual amount of publicity the case had garnered, the jurors were ordered to be sequestered – the first time in the history of American jurisprudence that such a restriction was ordered. The trial proceedings began on January 23, 1907, and the jury went into deliberation on April 11. After 47 hours, the jurors emerged deadlocked. Seven had voted guilty, and five voted not guilty. Thaw was outraged that the jurors had not recognized it as the act, as he saw it, of one chivalrous man defending innocent womanhood.


title: Evelyn Nesbit

text: The second trial took place from January 1908 through February 1, 1908. At the second trial, Thaw again pleaded temporary insanity. Thaw was found not guilty, on the ground of insanity at the time of the commission of his act. He was sentenced to involuntary commitment for life in the Matteawan State Hospital for the Criminally Insane in Beacon, New York. His wealth allowed him to arrange accommodations for his comfort and be granted privileges not given to the general Matteawan population. Immediately after his confinement, Thaw marshaled the forces of a legal team charged with the mission of having him declared sane; the effort took seven years. The prolonged legal procedures compelled his escape from Matteawan and flight to Canada in 1913; he was extradited to the U.S., but in 1915 was released from custody after being judged sane.


title: Evelyn Nesbit

text: Nesbit gave birth to a son, Russell William Thaw, on October 25, 1910, in Berlin, Germany. She always maintained that her son was Thaw's biological child, conceived during a conjugal visit to Thaw while he was confined at Matteawan, although Thaw denied paternity throughout his life.


title: Evelyn Nesbit

text: In 1911, Nesbit reconciled with her mother, who took on the role of caregiver for the child while Nesbit sought out opportunities to support herself and her son. Russell appeared with his mother in at least six films: "Threads of Destiny" (1914), "Redemption" (1917), "Her Mistake" (1918), "The Woman Who Gave" (1918), "I Want to Forget" (1918), and "The Hidden Woman" (1922). He became an accomplished pilot, placing third in the 1935 Bendix Trophy race from Los Angeles to Cleveland, ahead of Amelia Earhart in fifth place.


title: Evelyn Nesbit

text: Throughout the prolonged court proceedings, Nesbit had received financial support from the Thaws. These payments, made to her through the family's attorneys, had been inconsistent and far from generous. After the close of the second trial, the Thaws virtually abandoned her, cutting off all funds. Her grandson, Russell Thaw Jr., recounted a piece of family lore in a 2005 interview with the "Los Angeles Times": purportedly, she had received $25,000 from the Thaws after the culmination of the trials. To spite them, she then donated the money to anarchist Emma Goldman, who subsequently turned it over to investigative journalist and political activist John Reed. Nesbit was left to her own resources to provide for herself. She found modest success working in vaudeville and on the silent screen. In 1914, she appeared in "Threads of Destiny", produced at the Betzwood studios of film producer Siegmund Lubin.


title: Evelyn Nesbit

text: Nesbit divorced Thaw in 1915. As soon as the divorce was finalized, she married her dance partner, Jack Clifford in 1916. The announcement was front page news. Beginning in 1913, the couple had toured with an extremely successful stage act. In August 1913, she gave a dance performance at New York City's Victoria Theater, reported as her first performance on the New York City stage since 1904. Despite what one reviewer called an “indifferent vaudeville exhibition,” in November 1913, they packed the house at Chicago’s Auditorium Theater, drawing an overall audience of 7,400 at the venue, turning away hundreds. Their marriage did not fare as well. Clifford eventually found his wife's notoriety an insurmountable issue, with his own identity subsumed by that of "Mr. Evelyn Nesbit". He left her in 1918. After years of legal battles and accusations of infidelity, their divorce was finalized in 1933. Her long-term friend and employer, Dan Blanco, supported her in court. A well-known Chicago nightclub proprietor, Blanco helped engineer Nesbit’s cabaret comeback in the 1920s, first at Chicago’s Moulin Rouge and later at his own Club Alabam.


title: Evelyn Nesbit

text: In 1921, Nesbit briefly became the proprietor of a tearoom called The Evelyn Nesbit Specialty Food Shop, located in the West 50s in Manhattan, which may have doubled as a speakeasy. Through the 1920s and well into the 1930s, she struggled with chronic financial problems, alcoholism, and morphine addiction.


title: Evelyn Nesbit

text: Following her years in vaudeville, Nesbit transitioned to playing clubs and cabarets throughout the East, South, and Midwest. She briefly lent her name to several, including the Evelyn Nesbit Club (Atlantic City) and Chez Evelyn (Manhattan). Fond of Chicago audiences, she frequently played Rush Street’s Club Alabam.


title: Evelyn Nesbit

text: During the 1930s, she worked in Panama and added burlesque to her repertoire. In 1939, while sharing the bill with strippers, the then-53-year-old Nesbit told a "New York Times" reporter: "I wish I were a strip-teaser. I wouldn't have to bother with so many clothes."


title: Evelyn Nesbit

text: On New Year’s Eve 1925, Nesbit concluded a six-week engagement at Chicago’s Moulin Rouge and was scheduled to begin a booking in Miami, Florida, on January 2, 1926. Instead of heading to Florida, she went on a bender and attempted suicide by swallowing disinfectant. For days, headlines across the country once again turned Nesbit’s tragic life into front-page news. Later, doctors stated that Nesbit might have died if her stomach had not been full of gin.


title: Evelyn Nesbit

text: Nesbit and Harry K. Thaw continued to fascinate the public and the press speculated about the status of their relationship. Following her suicide attempt, one newspaper headline on January 8, 1926, said: "Thaw to Visit Chicago: Reconciliation Rumor". In an interview, Thaw said that he had been paying Nesbit $10 a day through an attorney, as a "token of pleasant memories of the past when we were happy". In June 1926, they were photographed together. On good terms, in 1927, Thaw attended the opening of Nesbit’s Manhattan café, Chez Evelyn and, in 1929, rumors flew that the couple intended to remarry and that Thaw had purchased an Atlantic City bungalow for Nesbit. When Thaw died in 1947, he bequeathed $10,000 to Nesbit from an estate valued at over $1 million.


title: Evelyn Nesbit

text: On June 5, 1945, Nesbit became national news yet again when she was questioned about the murder of Albert Langford, the husband of her friend, Marion Langford. The man was allegedly slain by one of two men who forcibly entered their Manhattan apartment. Nesbit had a strong alibi for the night of the murder and it was never suggested that she was in any way connected with the crime. Rather, her friendship with the Langfords became just another opportunity to use her name to sell newspapers. The case remains unsolved.


title: Evelyn Nesbit

text: Following Harry K. Thaw’s death in 1947, Nesbit left her home in New York City to settle in California. Her son, Russell W. Thaw, lived in West Los Angeles. She chose to live downtown in a Bohemian neighborhood located just north of Bunker Hill. There she pursued a long-standing interest in sculpting, studying at the Grant Beach School of Arts and Crafts. Following graduation in 1952, she taught classes in sculpting and ceramics.


title: Evelyn Nesbit

text: In the summer of 1955, Nesbit served as the technical adviser on the movie "The Girl in the Red Velvet Swing" (1955), for which she was paid $10,000. The movie recounts her early life and White's murder, blending fiction with fact. While working on the film, Nesbit collapsed from exhaustion.


title: Evelyn Nesbit

text: Nesbit died in a nursing home in Santa Monica, California, on January 17, 1967, at the age of 82. She had been a resident there for more than a year. She was buried at Holy Cross Cemetery in Culver City, California.


title: DNA sequencing

text: DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine. The advent of rapid DNA sequencing methods has greatly accelerated biological and medical research and discovery.


title: DNA sequencing

text: Knowledge of DNA sequences has become indispensable for basic biological research, DNA Genographic Projects and in numerous applied fields such as medical diagnosis, biotechnology, forensic biology, virology and biological systematics. Comparing healthy and mutated DNA sequences can diagnose different diseases including various cancers, characterize antibody repertoire, and can be used to guide patient treatment. Having a quick way to sequence DNA allows for faster and more individualized medical care to be administered, and for more organisms to be identified and cataloged.


title: DNA sequencing

text: The rapid speed of sequencing attained with modern DNA sequencing technology has been instrumental in the sequencing of complete DNA sequences, or genomes, of numerous types and species of life, including the human genome and other complete DNA sequences of many animal, plant, and microbial species.


title: DNA sequencing

text: The first DNA sequences were obtained in the early 1970s by academic researchers using laborious methods based on two-dimensional chromatography. Following the development of fluorescence-based sequencing methods with a DNA sequencer, DNA sequencing has become easier and orders of magnitude faster.


title: DNA sequencing

text: DNA sequencing may be used to determine the sequence of individual genes, larger genetic regions (i.e. clusters of genes or operons), full chromosomes, or entire genomes of any organism. DNA sequencing is also the most efficient way to indirectly sequence RNA or proteins (via their open reading frames). In fact, DNA sequencing has become a key technology in many areas of biology and other sciences such as medicine, forensics, and anthropology.


title: DNA sequencing

text: Sequencing is used in molecular biology to study genomes and the proteins they encode. Information obtained using sequencing allows researchers to identify changes in genes and noncoding DNA (including regulatory sequences), associations with diseases and phenotypes, and identify potential drug targets.


title: DNA sequencing

text: Since DNA is an informative macromolecule in terms of transmission from one generation to another, DNA sequencing is used in evolutionary biology to study how different organisms are related and how they evolved. In February 2021, scientists reported, for the first time, the sequencing of DNA from animal remains, a mammoth in this instance, over a million years old, the oldest DNA sequenced to date.


title: DNA sequencing

text: The field of metagenomics involves identification of organisms present in a body of water, sewage, dirt, debris filtered from the air, or swab samples from organisms. Knowing which organisms are present in a particular environment is critical to research in ecology, epidemiology, microbiology, and other fields. Sequencing enables researchers to determine which types of microbes may be present in a microbiome, for example.


title: DNA sequencing

text: As most viruses are too small to be seen by a light microscope, sequencing is one of the main tools in virology to identify and study the virus. Viral genomes can be based in DNA or RNA. RNA viruses are more time-sensitive for genome sequencing, as they degrade faster in clinical samples. Traditional Sanger sequencing and next-generation sequencing are used to sequence viruses in basic and clinical research, as well as for the diagnosis of emerging viral infections, molecular epidemiology of viral pathogens, and drug-resistance testing. There are more than 2.3 million unique viral sequences in GenBank. Recently, NGS has surpassed traditional Sanger as the most popular approach for generating viral genomes.


title: DNA sequencing

text: During the 1990 avian influenza outbreak, viral sequencing determined that the influenza sub-type originated through reassortment between quail and poultry. This led to legislation in Hong Kong that prohibited selling live quail and poultry together at market. Viral sequencing can also be used to estimate when a viral outbreak began by using a molecular clock technique.


title: DNA sequencing

text: Medical technicians may sequence genes (or, theoretically, full genomes) from patients to determine if there is risk of genetic diseases. This is a form of genetic testing, though some genetic tests may not involve DNA sequencing.


title: DNA sequencing

text: DNA sequencing is also being increasingly used to diagnose and treat rare diseases. As more and more genes are identified that cause rare genetic diseases, molecular diagnoses for patients becomes more mainstream. DNA sequencing allows clinicians to identify genetic diseases, improve disease management, provide reproductive counseling, and more effective therapies.


title: DNA sequencing

text: Also, DNA sequencing may be useful for determining a specific bacteria, to allow for more precise antibiotics treatments, hereby reducing the risk of creating antimicrobial resistance in bacteria populations.


title: DNA sequencing

text: DNA sequencing may be used along with DNA profiling methods for forensic identification and paternity testing. DNA testing has evolved tremendously in the last few decades to ultimately link a DNA print to what is under investigation. The DNA patterns in fingerprint, saliva, hair follicles, etc. uniquely separate each living organism from another. Testing DNA is a technique which can detect specific genomes in a DNA strand to produce a unique and individualized pattern.


title: DNA sequencing

text: The canonical structure of DNA has four bases: thymine (T), adenine (A), cytosine (C), and guanine (G). DNA sequencing is the determination of the physical order of these bases in a molecule of DNA. However, there are many other bases that may be present in a molecule. In some viruses (specifically, bacteriophage), cytosine may be replaced by hydroxy methyl or hydroxy methyl glucose cytosine. In mammalian DNA, variant bases with methyl groups or phosphosulfate may be found. Depending on the sequencing technique, a particular modification, e.g., the 5mC (5 methyl cytosine) common in humans, may or may not be detected.


title: DNA sequencing

text: Deoxyribonucleic acid (DNA) was first discovered and isolated by Friedrich Miescher in 1869, but it remained under-studied for many decades because proteins, rather than DNA, were thought to hold the genetic blueprint to life. This situation changed after 1944 as a result of some experiments by Oswald Avery, Colin MacLeod, and Maclyn McCarty demonstrating that purified DNA could change one strain of bacteria into another. This was the first time that DNA was shown capable of transforming the properties of cells.


title: DNA sequencing

text: In 1953, James Watson and Francis Crick put forward their double-helix model of DNA, based on crystallized X-ray structures being studied by Rosalind Franklin. According to the model, DNA is composed of two strands of nucleotides coiled around each other, linked together by hydrogen bonds and running in opposite directions. Each strand is composed of four complementary nucleotides – adenine (A), cytosine (C), guanine (G) and thymine (T) – with an A on one strand always paired with T on the other, and C always paired with G. They proposed that such a structure allowed each strand to be used to reconstruct the other, an idea central to the passing on of hereditary information between generations.


title: DNA sequencing

text: The foundation for sequencing proteins was first laid by the work of Frederick Sanger who by 1955 had completed the sequence of all the amino acids in insulin, a small protein secreted by the pancreas. This provided the first conclusive evidence that proteins were chemical entities with a specific molecular pattern rather than a random mixture of material suspended in fluid. Sanger's success in sequencing insulin spurred on x-ray crystallographers, including Watson and Crick, who by now were trying to understand how DNA directed the formation of proteins within a cell. Soon after attending a series of lectures given by Frederick Sanger in October 1954, Crick began developing a theory which argued that the arrangement of nucleotides in DNA determined the sequence of amino acids in proteins, which in turn helped determine the function of a protein. He published this theory in 1958.


title: DNA sequencing

text: RNA sequencing was one of the earliest forms of nucleotide sequencing. The major landmark of RNA sequencing is the sequence of the first complete gene and the complete genome of Bacteriophage MS2, identified and published by Walter Fiers and his coworkers at the University of Ghent (Ghent, Belgium), in 1972 and 1976. Traditional RNA sequencing methods require the creation of a cDNA molecule which must be sequenced.


title: DNA sequencing

text: The first method for determining DNA sequences involved a location-specific primer extension strategy established by Ray Wu at Cornell University in 1970. DNA polymerase catalysis and specific nucleotide labeling, both of which figure prominently in current sequencing schemes, were used to sequence the cohesive ends of lambda phage DNA. Between 1970 and 1973, Wu, R Padmanabhan and colleagues demonstrated that this method can be employed to determine any DNA sequence using synthetic location-specific primers. Frederick Sanger then adopted this primer-extension strategy to develop more rapid DNA sequencing methods at the MRC Centre, Cambridge, UK and published a method for "DNA sequencing with chain-terminating inhibitors" in 1977. Walter Gilbert and Allan Maxam at Harvard also developed sequencing methods, including one for "DNA sequencing by chemical degradation". In 1973, Gilbert and Maxam reported the sequence of 24 basepairs using a method known as wandering-spot analysis. Advancements in sequencing were aided by the concurrent development of recombinant DNA technology, allowing DNA samples to be isolated from sources other than viruses.


title: DNA sequencing

text: The first full DNA genome to be sequenced was that of bacteriophage φX174 in 1977. Medical Research Council scientists deciphered the complete DNA sequence of the Epstein-Barr virus in 1984, finding it contained 172,282 nucleotides. Completion of the sequence marked a significant turning point in DNA sequencing because it was achieved with no prior genetic profile knowledge of the virus.


title: DNA sequencing

text: A non-radioactive method for transferring the DNA molecules of sequencing reaction mixtures onto an immobilizing matrix during electrophoresis was developed by Herbert Pohl and co-workers in the early 1980s. Followed by the commercialization of the DNA sequencer "Direct-Blotting-Electrophoresis-System GATC 1500" by GATC Biotech, which was intensively used in the framework of the EU genome-sequencing programme, the complete DNA sequence of the yeast "Saccharomyces cerevisiae" chromosome II. Leroy E. Hood's laboratory at the California Institute of Technology announced the first semi-automated DNA sequencing machine in 1986. This was followed by Applied Biosystems' marketing of the first fully automated sequencing machine, the ABI 370, in 1987 and by Dupont's Genesis 2000 which used a novel fluorescent labeling technique enabling all four dideoxynucleotides to be identified in a single lane. By 1990, the U.S. National Institutes of Health (NIH) had begun large-scale sequencing trials on "Mycoplasma capricolum", "Escherichia coli", "Caenorhabditis elegans", and "Saccharomyces cerevisiae" at a cost of US$0.75 per base. Meanwhile, sequencing of human cDNA sequences called expressed sequence tags began in Craig Venter's lab, an attempt to capture the coding fraction of the human genome. In 1995, Venter, Hamilton Smith, and colleagues at The Institute for Genomic Research (TIGR) published the first complete genome of a free-living organism, the bacterium "Haemophilus influenzae". The circular chromosome contains 1,830,137 bases and its publication in the journal Science marked the first published use of whole-genome shotgun sequencing, eliminating the need for initial mapping efforts.


title: DNA sequencing

text: Several new methods for DNA sequencing were developed in the mid to late 1990s and were implemented in commercial DNA sequencers by 2000. Together these were called the "next-generation" or "second-generation" sequencing (NGS) methods, in order to distinguish them from the earlier methods, including Sanger sequencing. In contrast to the first generation of sequencing, NGS technology is typically characterized by being highly scalable, allowing the entire genome to be sequenced at once. Usually, this is accomplished by fragmenting the genome into small pieces, randomly sampling for a fragment, and sequencing it using one of a variety of technologies, such as those described below. An entire genome is possible because multiple fragments are sequenced at once (giving it the name "massively parallel" sequencing) in an automated process.


title: DNA sequencing

text: NGS technology has tremendously empowered researchers to look for insights into health, anthropologists to investigate human origins, and is catalyzing the "Personalized Medicine" movement. However, it has also opened the door to more room for error. There are many software tools to carry out the computational analysis of NGS data, often compiled at online platforms such as CSI NGS Portal, each with its own algorithm. Even the parameters within one software package can change the outcome of the analysis. In addition, the large quantities of data produced by DNA sequencing have also required development of new methods and programs for sequence analysis. Several efforts to develop standards in the NGS field have been attempted to address these challenges, most of which have been small-scale efforts arising from individual labs. Most recently, a large, organized, FDA-funded effort has culminated in the BioCompute standard.


title: DNA sequencing

text: On 26 October 1990, Roger Tsien, Pepi Ross, Margaret Fahnestock and Allan J Johnston filed a patent describing stepwise ("base-by-base") sequencing with removable 3' blockers on DNA arrays (blots and single DNA molecules).


title: DNA sequencing

text: In 1996, Pål Nyrén and his student Mostafa Ronaghi at the Royal Institute of Technology in Stockholm published their method of pyrosequencing.


title: DNA sequencing

text: On 1 April 1997, Pascal Mayer and Laurent Farinelli submitted patents to the World Intellectual Property Organization describing DNA colony sequencing. The DNA sample preparation and random surface-polymerase chain reaction (PCR) arraying methods described in this patent, coupled to Roger Tsien et al.'s "base-by-base" sequencing method, is now implemented in Illumina's Hi-Seq genome sequencers.


title: DNA sequencing

text: In 1998, Phil Green and Brent Ewing of the University of Washington described their phred quality score for sequencer data analysis, a landmark analysis technique that gained widespread adoption, and which is still the most common metric for assessing the accuracy of a sequencing platform.


title: DNA sequencing

text: Lynx Therapeutics published and marketed massively parallel signature sequencing (MPSS), in 2000. This method incorporated a parallelized, adapter/ligation-mediated, bead-based sequencing technology and served as the first commercially available "next-generation" sequencing method, though no DNA sequencers were sold to independent laboratories.


title: DNA sequencing

text: Allan Maxam and Walter Gilbert published a DNA sequencing method in 1977 based on chemical modification of DNA and subsequent cleavage at specific bases. Also known as chemical sequencing, this method allowed purified samples of double-stranded DNA to be used without further cloning. This method's use of radioactive labeling and its technical complexity discouraged extensive use after refinements in the Sanger methods had been made.


title: DNA sequencing

text: Maxam-Gilbert sequencing requires radioactive labeling at one 5' end of the DNA and purification of the DNA fragment to be sequenced. Chemical treatment then generates breaks at a small proportion of one or two of the four nucleotide bases in each of four reactions (G, A+G, C, C+T). The concentration of the modifying chemicals is controlled to introduce on average one modification per DNA molecule. Thus a series of labeled fragments is generated, from the radiolabeled end to the first "cut" site in each molecule. The fragments in the four reactions are electrophoresed side by side in denaturing acrylamide gels for size separation. To visualize the fragments, the gel is exposed to X-ray film for autoradiography, yielding a series of dark bands each corresponding to a radiolabeled DNA fragment, from which the sequence may be inferred.


title: DNA sequencing

text: The chain-termination method developed by Frederick Sanger and coworkers in 1977 soon became the method of choice, owing to its relative ease and reliability. When invented, the chain-terminator method used fewer toxic chemicals and lower amounts of radioactivity than the Maxam and Gilbert method. Because of its comparative ease, the Sanger method was soon automated and was the method used in the first generation of DNA sequencers.


title: DNA sequencing

text: Sanger sequencing is the method which prevailed from the 1980s until the mid-2000s. Over that period, great advances were made in the technique, such as fluorescent labelling, capillary electrophoresis, and general automation. These developments allowed much more efficient sequencing, leading to lower costs. The Sanger method, in mass production form, is the technology which produced the first human genome in 2001, ushering in the age of genomics. However, later in the decade, radically different approaches reached the market, bringing the cost per genome down from $100 million in 2001 to $10,000 in 2011.


title: DNA sequencing

text: Large-scale sequencing often aims at sequencing very long DNA pieces, such as whole chromosomes, although large-scale sequencing can also be used to generate very large numbers of short sequences, such as found in phage display. For longer targets such as chromosomes, common approaches consist of cutting (with restriction enzymes) or shearing (with mechanical forces) large DNA fragments into shorter DNA fragments. The fragmented DNA may then be cloned into a DNA vector and amplified in a bacterial host such as "Escherichia coli". Short DNA fragments purified from individual bacterial colonies are individually sequenced and assembled electronically into one long, contiguous sequence. Studies have shown that adding a size selection step to collect DNA fragments of uniform size can improve sequencing efficiency and accuracy of the genome assembly. In these studies, automated sizing has proven to be more reproducible and precise than manual gel sizing.


title: DNA sequencing

text: The term ""de novo" sequencing" specifically refers to methods used to determine the sequence of DNA with no previously known sequence. "De novo" translates from Latin as "from the beginning". Gaps in the assembled sequence may be filled by primer walking. The different strategies have different tradeoffs in speed and accuracy; shotgun methods are often used for sequencing large genomes, but its assembly is complex and difficult, particularly with sequence repeats often causing gaps in genome assembly.


title: DNA sequencing

text: Most sequencing approaches use an "in vitro" cloning step to amplify individual DNA molecules, because their molecular detection methods are not sensitive enough for single molecule sequencing. Emulsion PCR isolates individual DNA molecules along with primer-coated beads in aqueous droplets within an oil phase. A polymerase chain reaction (PCR) then coats each bead with clonal copies of the DNA molecule followed by immobilization for later sequencing. Emulsion PCR is used in the methods developed by Marguilis et al. (commercialized by 454 Life Sciences), Shendure and Porreca et al. (also known as "polony sequencing") and SOLiD sequencing, (developed by Agencourt, later Applied Biosystems, now Life Technologies). Emulsion PCR is also used in the GemCode and Chromium platforms developed by 10x Genomics.


title: DNA sequencing

text: Shotgun sequencing is a sequencing method designed for analysis of DNA sequences longer than 1000 base pairs, up to and including entire chromosomes. This method requires the target DNA to be broken into random fragments. After sequencing individual fragments using the chain termination method, the sequences can be reassembled on the basis of their overlapping regions.


title: DNA sequencing

text: High-throughput sequencing, which includes next-generation "short-read" and third-generation "long-read" sequencing methods, applies to exome sequencing, genome sequencing, genome resequencing, transcriptome profiling (RNA-Seq), DNA-protein interactions (ChIP-sequencing), and epigenome characterization.


title: DNA sequencing

text: The high demand for low-cost sequencing has driven the development of high-throughput sequencing technologies that parallelize the sequencing process, producing thousands or millions of sequences concurrently. High-throughput sequencing technologies are intended to lower the cost of DNA sequencing beyond what is possible with standard dye-terminator methods. In ultra-high-throughput sequencing as many as 500,000 sequencing-by-synthesis operations may be run in parallel. Such technologies led to the ability to sequence an entire human genome in as little as one day. , corporate leaders in the development of high-throughput sequencing products included Illumina, Qiagen and ThermoFisher Scientific.


title: DNA sequencing

text: SMRT sequencing is based on the sequencing by synthesis approach. The DNA is synthesized in zero-mode wave-guides (ZMWs) – small well-like containers with the capturing tools located at the bottom of the well. The sequencing is performed with use of unmodified polymerase (attached to the ZMW bottom) and fluorescently labelled nucleotides flowing freely in the solution. The wells are constructed in a way that only the fluorescence occurring by the bottom of the well is detected. The fluorescent label is detached from the nucleotide upon its incorporation into the DNA strand, leaving an unmodified DNA strand. According to Pacific Biosciences (PacBio), the SMRT technology developer, this methodology allows detection of nucleotide modifications (such as cytosine methylation). This happens through the observation of polymerase kinetics. This approach allows reads of 20,000 nucleotides or more, with average read lengths of 5 kilobases. In 2015, Pacific Biosciences announced the launch of a new sequencing instrument called the Sequel System, with 1 million ZMWs compared to 150,000 ZMWs in the PacBio RS II instrument. SMRT sequencing is referred to as "third-generation" or "long-read" sequencing.


title: DNA sequencing

text: The DNA passing through the nanopore changes its ion current. This change is dependent on the shape, size and length of the DNA sequence. Each type of the nucleotide blocks the ion flow through the pore for a different period of time. The method does not require modified nucleotides and is performed in real time. Nanopore sequencing is referred to as "third-generation" or "long-read" sequencing, along with SMRT sequencing.


title: DNA sequencing

text: Early industrial research into this method was based on a technique called 'exonuclease sequencing', where the readout of electrical signals occurred as nucleotides passed by alpha(α)-hemolysin pores covalently bound with cyclodextrin. However the subsequent commercial method, 'strand sequencing', sequenced DNA bases in an intact strand.


title: DNA sequencing

text: Two main areas of nanopore sequencing in development are solid state nanopore sequencing, and protein based nanopore sequencing. Protein nanopore sequencing utilizes membrane protein complexes such as α-hemolysin, MspA ("Mycobacterium smegmatis" Porin A) or CssG, which show great promise given their ability to distinguish between individual and groups of nucleotides. In contrast, solid-state nanopore sequencing utilizes synthetic materials such as silicon nitride and aluminum oxide and it is preferred for its superior mechanical ability and thermal and chemical stability. The fabrication method is essential for this type of sequencing given that the nanopore array can contain hundreds of pores with diameters smaller than eight nanometers.


title: DNA sequencing

text: The concept originated from the idea that single stranded DNA or RNA molecules can be electrophoretically driven in a strict linear sequence through a biological pore that can be less than eight nanometers, and can be detected given that the molecules release an ionic current while moving through the pore. The pore contains a detection region capable of recognizing different bases, with each base generating various time specific signals corresponding to the sequence of bases as they cross the pore which are then evaluated. Precise control over the DNA transport through the pore is crucial for success. Various enzymes such as exonucleases and polymerases have been used to moderate this process by positioning them near the pore's entrance.


title: DNA sequencing

text: The first of the high-throughput sequencing technologies, massively parallel signature sequencing (or MPSS), was developed in the 1990s at Lynx Therapeutics, a company founded in 1992 by Sydney Brenner and Sam Eletr. MPSS was a bead-based method that used a complex approach of adapter ligation followed by adapter decoding, reading the sequence in increments of four nucleotides. This method made it susceptible to sequence-specific bias or loss of specific sequences. Because the technology was so complex, MPSS was only performed 'in-house' by Lynx Therapeutics and no DNA sequencing machines were sold to independent laboratories. Lynx Therapeutics merged with Solexa (later acquired by Illumina) in 2004, leading to the development of sequencing-by-synthesis, a simpler approach acquired from Manteia Predictive Medicine, which rendered MPSS obsolete. However, the essential properties of the MPSS output were typical of later high-throughput data types, including hundreds of thousands of short DNA sequences. In the case of MPSS, these were typically used for sequencing cDNA for measurements of gene expression levels.


title: DNA sequencing

text: The polony sequencing method, developed in the laboratory of George M. Church at Harvard, was among the first high-throughput sequencing systems and was used to sequence a full "E. coli" genome in 2005. It combined an in vitro paired-tag library with emulsion PCR, an automated microscope, and ligation-based sequencing chemistry to sequence an "E. coli" genome at an accuracy of >99.9999% and a cost approximately 1/9 that of Sanger sequencing. The technology was licensed to Agencourt Biosciences, subsequently spun out into Agencourt Personal Genomics, and eventually incorporated into the Applied Biosystems SOLiD platform. Applied Biosystems was later acquired by Life Technologies, now part of Thermo Fisher Scientific.


title: DNA sequencing

text: A parallelized version of pyrosequencing was developed by 454 Life Sciences, which has since been acquired by Roche Diagnostics. The method amplifies DNA inside water droplets in an oil solution (emulsion PCR), with each droplet containing a single DNA template attached to a single primer-coated bead that then forms a clonal colony. The sequencing machine contains many picoliter-volume wells each containing a single bead and sequencing enzymes. Pyrosequencing uses luciferase to generate light for detection of the individual nucleotides added to the nascent DNA, and the combined data are used to generate sequence reads. This technology provides intermediate read length and price per base compared to Sanger sequencing on one end and Solexa and SOLiD on the other.


title: DNA sequencing

text: Solexa, now part of Illumina, was founded by Shankar Balasubramanian and David Klenerman in 1998, and developed a sequencing method based on reversible dye-terminators technology, and engineered polymerases. The reversible terminated chemistry concept was invented by Bruno Canard and Simon Sarfati at the Pasteur Institute in Paris. It was developed internally at Solexa by those named on the relevant patents. In 2004, Solexa acquired the company Manteia Predictive Medicine in order to gain a massively parallel sequencing technology invented in 1997 by Pascal Mayer and Laurent Farinelli. It is based on "DNA clusters" or "DNA colonies", which involves the clonal amplification of DNA on a surface. The cluster technology was co-acquired with Lynx Therapeutics of California. Solexa Ltd. later merged with Lynx to form Solexa Inc.


title: DNA sequencing

text: In this method, DNA molecules and primers are first attached on a slide or flow cell and amplified with polymerase so that local clonal DNA colonies, later coined "DNA clusters", are formed. To determine the sequence, four types of reversible terminator bases (RT-bases) are added and non-incorporated nucleotides are washed away. A camera takes images of the fluorescently labeled nucleotides. Then the dye, along with the terminal 3' blocker, is chemically removed from the DNA, allowing for the next cycle to begin. Unlike pyrosequencing, the DNA chains are extended one nucleotide at a time and image acquisition can be performed at a delayed moment, allowing for very large arrays of DNA colonies to be captured by sequential images taken from a single camera.


title: DNA sequencing

text: Decoupling the enzymatic reaction and the image capture allows for optimal throughput and theoretically unlimited sequencing capacity. With an optimal configuration, the ultimately reachable instrument throughput is thus dictated solely by the analog-to-digital conversion rate of the camera, multiplied by the number of cameras and divided by the number of pixels per DNA colony required for visualizing them optimally (approximately 10 pixels/colony). In 2012, with cameras operating at more than 10 MHz A/D conversion rates and available optics, fluidics and enzymatics, throughput can be multiples of 1 million nucleotides/second, corresponding roughly to 1 human genome equivalent at 1x coverage per hour per instrument, and 1 human genome re-sequenced (at approx. 30x) per day per instrument (equipped with a single camera).


title: DNA sequencing

text: This method is an upgraded modification to combinatorial probe anchor ligation technology (cPAL) described by Complete Genomics which has since become part of Chinese genomics company BGI in 2013. The two companies have refined the technology to allow for longer read lengths, reaction time reductions and faster time to results. In addition, data are now generated as contiguous full-length reads in the standard FASTQ file format and can be used as-is in most short-read-based bioinformatics analysis pipelines.


title: DNA sequencing

text: The two technologies that form the basis for this high-throughput sequencing technology are DNA nanoballs (DNB) and patterned arrays for nanoball attachment to a solid surface. DNA nanoballs are simply formed by denaturing double stranded, adapter ligated libraries and ligating the forward strand only to a splint oligonucleotide to form a ssDNA circle. Faithful copies of the circles containing the DNA insert are produced utilizing Rolling Circle Amplification that generates approximately 300–500 copies. The long strand of ssDNA folds upon itself to produce a three-dimensional nanoball structure that is approximately 220 nm in diameter. Making DNBs replaces the need to generate PCR copies of the library on the flow cell and as such can remove large proportions of duplicate reads, adapter-adapter ligations and PCR induced errors.


title: DNA sequencing

text: The patterned array of positively charged spots is fabricated through photolithography and etching techniques followed by chemical modification to generate a sequencing flow cell. Each spot on the flow cell is approximately 250 nm in diameter, are separated by 700 nm (centre to centre) and allows easy attachment of a single negatively charged DNB to the flow cell and thus reducing under or over-clustering on the flow cell.


title: DNA sequencing

text: Sequencing is then performed by addition of an oligonucleotide probe that attaches in combination to specific sites within the DNB. The probe acts as an anchor that then allows one of four single reversibly inactivated, labelled nucleotides to bind after flowing across the flow cell. Unbound nucleotides are washed away before laser excitation of the attached labels then emit fluorescence and signal is captured by cameras that is converted to a digital output for base calling. The attached base has its terminator and label chemically cleaved at completion of the cycle. The cycle is repeated with another flow of free, labelled nucleotides across the flow cell to allow the next nucleotide to bind and have its signal captured. This process is completed a number of times (usually 50 to 300 times) to determine the sequence of the inserted piece of DNA at a rate of approximately 40 million nucleotides per second as of 2018.