
ekal-koshika anukraman

ekal-koshika anukraman agali peedhi kii anukraman praudyogikiyon key saath vyaktigat koshikaon seey anukram jaankaari kii jaanch kartaa haye, joe koshikiya antar kaa uchch sankalp aur isake sookshm vatavaran key sandarbh mein ack vyaktigat koshika key kaary kii behtar samajh pradaan kartaa haye. udaaharan key liye, cancer mein, vyaktigat koshikaon key dna koo anukramit karane seey koshikaon kii chhoti aabaadi dwara kiye gaye utparivartan key baare mein jaankaari mill sakati haye. vikas mein, vyaktigat koshikaon dwara vyakt aar. aine. aeye. kaa anukraman vibhinn koshika prakaron key astitva aur vyavhaar mein antardrishti dey sakata haye. sookshmajeev pranaliyon mein, ack hii prajaati kii aabaadi aanuvanshik roop seey clonel dikhaai dey sakati haye. phir bhee, aar. aine. aeye. yaa epigenetic sanshodhanon kii ekal-koshika anukraman koshika-seey-koshika parivartansheelta koo prakat curr sakata haye joe aabaadi koo badalate vatavaran mein jeevit rahane key liye teji seey anukoolit karane mein madad curr sakata haye.
prishthabhoomi.
ack vishisht maanav koshika mein lagbhag 2 x 3.3 billion aadhar jode dee. aine. aeye. aur 600 million aim. aar. aine. aeye. aadhar hotey hain. aamtaur parr, laakhon koshikaon key mishran kaa upayog dee. aine. aeye. yaa aar. aine. aeye. koo anukramit karane mein paaramparik tarikon jaise kii singer anukraman yaa roshni anukraman kaa upayog karke kiya jaataa haye. ekal koshika seey dee. aine. aeye. aur aar. aine. aeye. key gahan anukram dwara, koshikiya karyon kii vyaapak roop seey jaanch kii jaa sakati haye. vishisht agali peedhi key anukraman prayogon kii tarah, ekal-koshika anukraman protocol mein aam taur parr nimnalikhit charan hotey hainh ekal koshika kaa algaav, nuclic acid nishkarshan aur pravardhan, anukraman pustakalaya kii taiyari, anukraman aur jaiv soochanaatmak detta vishleshan. thoke mein koshikaon seey anukraman karane kii tulanaa mein ekal-koshika anukraman karna adhik chunautipurna haye. ekal koshika seey praarambhik samagri kii nyuntam matra ksharan, namuna haani aur sandushan koo anukraman detta kii gunavatta parr spasht prabhaav daalati haye. isake alawa, upayog kiye jaane vaale nuclic acid kii sankhya key picogram star key kaaran, ekal-koshika anukraman key namuna taiyar karane key dauran aksar bhaari pravardhan kii aavashyakta hoti haye, jisake parinamswarup asman coverage, shore aur anukraman detta kii galat matra hoti haye.
haal key takniki sudhaar ekal-koshika anukraman koo durgam prateet honey vaali samasyaon key ack samooh taka pahunchane key liye ack ashajanak upkaran banaate hain. udaaharan key liye, visham namoone, durlabh koshika prakaar, koshika vansh sambandh, shaaririk ootakon kaa mozekwad, sookshmajeevon kaa vishleshan jinhen samvardhit naheen kiya jaa sakata haye, aur roga vikas sabhi koo ekal-koshika anukraman key madhyam seey spasht kiya jaa sakata haye. prakriti prakaashan samooh dwara varsh 2013 kii vidhi key roop mein ekal-koshika anukraman kaa chayan kiya gayaa thaa.
genome (dee. aine. aeye.) anukraman.
ekal-koshika dna genome anukraman mein ack ekal koshika koo alag karna, poore genome yaa ruchi key kshetra koo badhaana, anukraman pustakalayon kaa nirmaan karna aur phir agali peedhi key dna anukraman (udaaharan key liye roshni, aayan dhaara, mgi) koo lagu karna shamil haye. samanya shareer vigyaan aur roga kaa adhyayan karane key liye standhari pranaliyon mein ekal-koshika dna anukraman vyaapak roop seey lagu kiya gayaa haye. ekal-koshika sankalp cancer key vikas yaa upchaar pratikriya mein aanuvanshik mozekwad yaa intra-tumour aanuvanshik vishamta kii bhoomikaon koo ujagar curr sakata haye. sookshmajeevon key sandarbh mein, ekal ekkoshikiya jeev seey ack genome koo ekal pravardhit genome (saig) key roop mein sandarbhit kiya jaataa haye. ekal-koshika dna anukraman mein pragati ney jatil sookshm jeevon mein maujood asanskrit procariotic prajaatiyon seey genomic detta ekatra karane mein saksham banaaya haye.  halanki saig kii visheshta cumm poornata aur mahatvapurna purvagrah haye, haal hii mein computational pragati ney samagr saig seey lagbhag poorna genome kii assembly hassil kii haye. sookshmajeevon seey praapt detta bhavishya mein samvardhan key liye prakriyaaen sthaapit curr sakata haye. ekal koshika ekal-koshika anukraman mein upayog kiye jaane vaale kuchh genome assembly upkaranon mein kudal, idba-uda, courtex aur hyda shamil hain.
vidhiyaan.
100 seey adhik vibhinn ekal-koshika omics vidhiyon kii ack soochi prakaashit kii gaee haye.
ekaadhik visthaapan pravardhan (aim. dee. aeye.) ack vyaapak roop seey upayog kii jaane vaali takneek haye, joe anukraman key liye jeevaanu seey microgram taka dee. aine. aeye. key femtogram koo pravardhit karane mein saksham banaati haye. aim. dee. aeye. pratikriyaon key liye aavashyak abhikarmakon mein shamil hainh yaadrichhik primer aur dee. aine. aeye. polymerase joe bacteriophase faaee29 seey hotey hain. 30 degree samtaap pratikriya mein, dee. aine. aeye. koo shamil abhikarmakon key saath pravardhit kiya jaataa haye. jaise hii bahulak nae taaron kaa nirmaan karate hain, ack taar visthaapan pratikriya hoti haye, joe pratyek template dna seey kaee pratiyon koo sanshleshit karati haye. usee samay, joe taar pahale seey vistaarit kiye gaye they, way visthaapit hoe jaayenge. aim. dee. aeye. utpaadon kii lambai lagbhag 12 key. bee. hoti haye aur isaki seema lagbhag 100 key. bee. taka hoti haye, jisase dee. aine. aeye. anukraman mein iska upayog kiya jaa sakata haye. 2017 mein, iss takneek mein ack badaa sudhaar, jise doubleu. jee. aeye.-aiks. kahaa jaataa haye, koo faaee29 polymerase key thermostable utparivarti kaa laabh uthaakar pesh kiya gayaa thaa, jisase vyaktigat koshikaon seey behtar genome recovery hui, vishesh roop seey uchch jee + see samagri vaale. aim. dee. aeye. koo ack atyadhik samanantar ekal-koshika poore genome pravardhan koo praapt karane key liye ack sookshm-drav bindu-adharit pranali mein bhee lagu kiya gayaa haye. dee. aine. aeye. pakadane aur pravardhan key liye ekal-koshikaon koo boondon mein samaahit karke, yeh vidhi paaramparik aim. dee. aeye. kii tulanaa mein cumm purvagrah aur unnat thruput pradaan karati haye.
ack anya samanya vidhi malback haye. aim. dee. aeye. mein kiya gayaa, yeh vidhi samtaap pravardhan key saath shuru hoti haye, lekin primer koo downstream pee. see. aar. pravardhan key liye ack "samanya" anukram key saath flank kiya jaataa haye. jaise-jaise praarambhik emplican utpann hotey hain, samanya anukram aatm-bandhan aur aage key pravardhan koo rokane key liye "loops" key gathan koo badhaava detaa haye. aim. dee. aeye. key vipareet, atyadhik shaakhaon vaalaa dee. aine. aeye. network naheen banata haye. isake bajaay, loop koo ack anya tapman chakr mein vikrit kiya jaataa haye jisase tukadon koo pee. see. aar. key saath pravardhit kiya jaa sakata haye. malback koo ack microfluidic upkaran mein bhee lagu kiya gayaa haye, lekin nanoliter boondon mein sankshepan seey pravardhan pradarshan mein kaafee sudhaar naheen huaa thaa.
aim. dee. aeye. aur malback kii tulanaa mein, aim. dee. aeye. key parinamswarup behtar genome coverage hotaa haye, lekin malback poore genome mein adhik samaan coverage pradaan kartaa haye. aim. dee. aeye. snaps kii pehchaan karane key liye adhik prabhaavi hoe sakata haye, jabaki malback koo coppy number sanskaranon kaa pataa lagaane key liye prathamikta dee jaati haye. ack sookshm drav upkaran key saath aim. dee. aeye. karane key dauran purvagrah aur sandushan koo ullekhaniya roop seey cumm curr detaa haye, malback mein shamil rasayan vigyaan behtar dakshata key liye samaan kshamata kaa pradarshan naheen kartaa haye. 
genomic sanrachanaatmak bhinnata kii khoj key liye vishesh roop seey upayukt ack vidhi ekal-koshika dna template strand anukraman (aeye.key.aeye. strand-seik). ekal-koshika tri-channel prasanskaran key siddhant kaa upayog karate hue, joe pathan-abhivinyas, pathan-gaharaai aur haplotype-charan key sanyukt modeling kaa upayog kartaa haye, strand-seik aakar mein shaaririk sanrachanaatmak bhinnata vargon ≤200kb key poorna spectrum kii khoj koo saksham banaata haye. strand-seik ekal koshikaon mein shaaririk aanuvanshik bhinnata vargon kii pehchaan key liye poore genome pravardhan adharit tarikon kii seemaon koo paar curr jaataa haye, kyonki yeh kalaakriti koo bulaane key liye padhane vaale chimmer key khilaf atisamvedansheel naheen haye (neeche diye gaye khand mein vistaar seey charcha kii gaee haye), aur drop out seey cumm prabhaavit hotaa haye. vidhi kaa chayan anukraman key lakshya parr nirbhar kartaa haye kyonki pratyek vidhi alag-alag laabh prastut karati haye.
seemaaen.
vyaktigat koshika genome key aim. dee. aeye. key parinamswarup atyadhik asman genome coverage hotaa haye.ee. template key vibhinn kshetron kaa saapeksh atee pratinidhitva aur cumm pratinidhitva, jisase kuchh anukramon kaa nuksan hotaa haye. iss prakriya key dou ghatak hainh aeye) yaadrichhik kshetron kaa yaadrichhik roop seey adhik aur cumm vistaar; aur bee) uchch% jee. see. kshetron key khilaf vyavasthit purvagrah. yaadrichhik ghatak koo usee koshika prakaar seey ekal-koshika aim. dee. aeye. pratikriyaon koo ekatra karke sambodhit kiya jaa sakata haye, jisamen fluorescent koo situ sankaran (machhali) aur/yaa anukraman key baad kii pushti mein niyojit kiya jaa sakata haye. uchch% jee. see. kshetron key khilaf aim. dee. aeye. key purvagrah koo thermostable polymerase kaa upayog karke sambodhit kiya jaa sakata haye, jaise kii doubleu. jee. aeye.-aiks. naamak prakriya mein.
ekal-nucleotide bahuroopata (aiss. aine. pee. aiss.), joe maanav genome mein aanuvanshik bhinnata kaa ack badaa hissa haye, aur pratilipi sankhya bhinnata (see. aine. vee.), ekal koshika anukraman mein samasyaaen paida karati haye, saath hii saath ekal koshika seey nikaale gaye dee. aine. aeye. kii seemit matra bhee. dee. aine. aeye. kii cumm matra key kaaran, dee. aine. aeye. kaa sateek vishleshan pravardhan key baad bhee samasyaaen paida kartaa haye kyonki coverage cumm hoti haye aur trutiyon key liye atisamvedansheel hoti haye. aim. dee. aeye. key saath, ausat genome coverage 80 pratishat seey cumm haye aur jinn aiss. aine. pee. koo anukraman paath dwara cover naheen kiya jaataa haye, unhein baahar curr diyaa jaayega. isake alawa, aim. dee. aeye. elil dropout kaa ack uchch anupaat dikhaata haye, vishamyugma key namoonon seey elil kaa pataa naheen lagaata haye. vibhinn aiss. aine. pee. algorithm vartamaan mein upayog mein hain lekin koee bhee ekal-koshika anukraman key liye vishisht naheen hain. cnv key saath mda bhee vaastavik cnv koo chhipaane vaale jhoothe cnv kii pehchaan karane kii samasya paida kartaa haye. ise hull karane key liye, jaba nakali see. aine. vee. seey pattern utpann kiye jaa sakate hain, tou algorithm iss shore kaa pataa lagaa sakate hain aur sahee roopon kaa utpaadan karane key liye ise mitaa sakate hain. 
strand-seik aanuvanshik sanskaran key liye poore genome pravardhan key aadhar parr vidhiyon kii seemaon koo paar kartaa haih choonki strand-seik koo cnv yaa pratilipi-santulit sanrachanaatmak sanskaran vargon kii seemaon (yaa breakpoint) koo paar karane key liye padhane (yaa padhane vaale jode) kii aavashyakta naheen hoti haye, yeh poore genome pravardhan parr adharit ekal-koshika vidhiyon kii samanya kalakritiyon key liye cumm atisamvedansheel haye, jisamen sanskaran breakpoint parr padhane aur padhane vaale chimera parr padhane key abhaav key kaaran sanskaran coll dropout shamil hain.strand-seik cumm seey cumm 200key. bee. aakar key sanrachanaatmak bhinnata vargon key poorna spectrum kii khoj kartaa haye, jisamen toot-sanlayan-pull chakr aur chromothrypsis ghatanaaen, saath hii santulit vyutkram, aur pratilipi-sankhya santulit yaa asantulit sthanantaran shamil hain."strand-seik dwara kiye gaye sanrachanaatmak sanskaran coll gunasutra-lambai haplotype dwara hull kiye jaate hain, joe atirikt sanskaran vishishtata pradaan kartaa haye. vartamaan seema key roop mein, strand-seik key liye bromodioxuridine (bradu) kaa upayog karke strand-vishisht labeling key liye vibhajit koshikaon kii aavashyakta hoti haye, aur vidhi aakar mein 200kb seey chhote roopon kaa pataa naheen lagaati haye, jaise kii mobile tatva sammilan.
anuprayog.
adhikansh vatavaran mein adhikansh sookshmajeevon koo samvardhit karane mein kathinaaee key kaaran sookshm jeev ekal koshika genomics key mukhya lakshyon mein seey hain. ekal-koshika genomics binaa kheti key sookshmajeev genome anukram praapt karane kaa ack shaktishali tarika haye. sookshmajeev paaristhitiki, vikas, saarvajanik swaasthya aur jaiv praudyogiki kshamata seey sambandhit prashnon kii ack vistrit shrinkhala koo sambodhit karane key liye iss drishtikon koo samudri, mitty, upasatah, jeev aur anya prakaar key sookshmajeevon parr vyaapak roop seey lagu kiya gayaa haye.
cancer anukraman bhee aiss. see. dee. aine. aeye. see. kyu. kaa ack ubharata huaa anuprayog haye. taaja yaa jamey hue tumour kaa vishleshan kiya jaa sakata haye aur poore genome dna drishtikon kaa upayog karke scan, snv aur punarvyavastha key sambandh mein kaafee achchi tarah seey vargeekrit kiya jaa sakata haye. cancer aiss. see. dee. aine. aeye. aiss. ee. kyu. vishesh roop seey samvardhit chikitsiya lakshyon jaise receptor tyrosine kinnez jeane (ee. jee. aff. aar., pee. dee. jee. aff. aar. aadi) mein maujood jatilata aur yaugik utparivartan kii gaharaai kii jaanch karane key liye upayogi haye.) jahan bulk tumour key paaramparik jansankhya-star key drishtikon tumour kii ekal koshikaon key bheetar inn utparivartan key saha-ghatana pattern koo hull karane mein saksham naheen hain. iss tarah kaa ativyaapi maarg sakriyan aur tumour koshika pratirodh kii atirek pradaan curr sakata haye.
dee. aine. aeye. methyllum anukraman.
ekal-koshika dna methyllum anukraman dna methylan kii matra nirdhaarit kartaa haye. prakriti mein honey vaale kaee gyaat prakaar key methylation hain, jinamen 5-methylcytocene (5 mc), 5-hydroxymethylsitiocytocyte (5 hmc), 6-methyladenine (6ma), aur 4 mc 4-methylcytocene (4 mc) shamil hain. eukariotes mein, vishesh roop seey jaanvaron mein, 5 mc genome key saath vyaapak haye aur pargamya tatvon koo dabaakar jeane abhivyakti koo niyantrit karane mein mahatvapurna bhoomika nibhaata haye. vyaktigat koshikaon mein 5 mc kaa anukraman yeh prakat curr sakata haye kii kaise ack ootak yaa aabaadi seey aanuvanshik roop seey samaan koshikaon mein epigenetic parivartan vibhinn phenotype vaali koshikaon koo janm dete hain.
vidhiyaan.
ekal koshikaon mein 5 mc kaa pataa lagaane aur anukraman karane mein bisulfite anukraman swarn maanak bunn gayaa haye. bisulfite key saath dee. aine. aeye. kaa upchaar cytosine avasheshon koo uracil mein parivartit kartaa haye, lekin 5-methylcytocene avasheshon koo aprabhaavit chhod detaa haye. isaliye, jiss dna kaa bisulfite key saath upchaar kiya gayaa haye, vaha kewal methylated cytosine koo hii barkarar rakhta haye. methyllum readout praapt karane key liye, bisulfite-upchaarit anukram koo ack aparivartit genome key saath sanrekhit kiya jaataa haye. 2014 mein ekal koshikaon mein poore genome bisulfite anukraman koo praapt kiya gayaa thaa. yeh vidhi vishisht prakriya seey judey dna key nuksan koo door karati haye, jahaan bisulfite vikhandan seey pahale anukraman adaptor jode jaate hain. isake bajaay, dee. aine. aeye. kaa upchaar karane aur bisulfite key saath vikhandit honey key baad adaptor jode jaate hain, jisase sabhi tukadon koo pee. see. aar. dwara pravardhit kiya jaa sakata haye. gahan anukraman kaa upayog karate hue, yeh vidhi pratyek koshika mein kull see. pee. jee. key ~40% koo pakadati haye. maujooda takneek key saath dee. aine. aeye. koo bisulfite upchaar seey pahale pravardhit naheen kiya jaa sakata haye, kyonki 5 mc key nishan polymerase dwara pratilipi naheen kiye jaayenge.
ekal-koshika cumm pratinidhitva bisulfite anukraman (scrubs) ack aur vidhi haye. yeh vidhi uchch see. pee. jee. samagri vaale genome key kshetron key liye samriddh karane key liye see. pee. jee. dweepon (see. jee. aaee. aiss.) parr samooh karane key liye methylated cytosine kii pravritti kaa laabh uthaati haye. yeh poore genome bisulfite anukraman kii tulanaa mein anukraman kii laagat koo cumm kartaa haye, lekin iss vidhi key coverage koo seemit kartaa haye. jaba aar. aar. bee. koo thoke namoonon parr lagu kiya jaataa haye, tou jeane pravartakon mein adhikansh see. pee. jee. saiton kaa pataa lagaaya jaataa haye, lekin jeane pravartakon mein site poore genome mein see. pee. jee. saiton kaa kewal 10 pratishat haye. ekal koshikaon mein, thoke namoone seey 40 pratishat see. pee. jee. saiton kaa pataa lagaaya jaataa haye. coverage badhaane key liye, iss vidhi koo ekal koshikaon key ack chhote pool parr bhee lagu kiya jaa sakata haye. 20 pool ekal koshikaon key namoone mein, thoke namoone seey 63 pratishat see. pee. jee. saiton kaa pataa lagaaya gayaa thaa. ekal koshikaon koo ikattha karna methyllum coverage koo badhaane kii ack ranneeti haye, lekin koshikaon kii aabaadi mein vishamta koo aspasht karane kii keemat parr.
seemaaen.
jabaki 5 mc kaa pataa lagaane key liye bisulfite anukraman sabse vyaapak roop seey upayog kiya jaane vaalaa drishtikon banaa huaa haye, rasayanik upchaar kathor aur tukade haye aur dna koo kharaab kartaa haye. yeh prabhaav taba badhata haye jaba thoke namoonon seey ekal koshikaon mein sthanantarit kiya jaataa haye. dee. aine. aeye. methylation kaa pataa lagaane key anya tarikon mein methylation-samvedansheel pratibandh enzyme shamil hain. pratibandh enzyme anya prakaar key methylation kaa pataa lagaane mein bhee saksham banaate hain, jaise kii dee. pee. aine. aaee. key saath 6ma. nanopore-adharit anukraman mool dna mein vikhandan yaa sanshodhan key binaa pratyaksh methylation anukraman key liye ack maarg bhee pradaan kartaa haye. nanopore anukraman kaa upayog bacteria key methylom koo anukramit karane key liye kiya gayaa haye, jinamen 6ma aur 4 mc (eukariotes mein 5 mc key vipareet) kaa prabhutva haye, lekin iss takneek koo abhee taka ekal koshikaon mein naheen badhaaya gayaa haye.
anuprayog.
aanuvanshik roop seey samaan koshikaon mein epigenetic antar kaa pataa lagaane key liye ekal-koshika dna methylation anukraman kaa vyaapak roop seey upayog kiya gayaa haye. inn vidhiyon koo unake vikas key dauran maanya karane key liye, ack mishrit aabaadi key ekal-koshika methyllum detta koo alag-alag koshika prakaron kii pehchaan karane key liye padaanukramit clustering dwara safaltapurvak vargeekrit kiya gayaa thaa. ack anya anuprayog praarambhik vikas mein pahale kuchh koshika vibhajanon key dauran ekal koshikaon kaa adhyayan curr rhaa haye taaki yeh samajha jaa sakey kii ekal bhroon seey vibhinn prakaar kii koshikaaen kaise nikalati hain. ekal-koshika sampoorn-genome bisulfite anukraman kaa upayog cancer mein durlabh lekin atyadhik sakriya koshika prakaron jaise parisanchari tumour koshikaon (ctc) kaa adhyayan karane key liye bhee kiya gayaa haye.
transposage-accessible chromatin sequencing (scatech-seik).
ekal koshika pargaman-sulabh chromatin anukraman genome mein chromatin kii pahunch kaa maanchitra banaata haye. ack transposage seedhe chromatin key khule kshetron mein anukraman adaptor daaltaa haye, jisase una kshetron koo pravardhit aur anukramit kiya jaa sakata haye.
pratilekh anukraman (scrana-seik).
microere aur bulk aar. aine. aeye.-seik jaisi maanak vidhiyaan koshikaon kii badee aabaadi seey aar. aine. aeye. abhivyakti kaa vishleshan karati hain. yeye maap mishrit-koshika aabaadi mein vyaktigat koshikaon key beach mahatvapurna antar koo aspasht curr sakate hain.
ekal-koshika aar. aine. aeye. anukraman (aiss. aar. aine. aeye.-seey. kyu.) vyaktigat koshikaon kii abhivyakti profile pradaan kartaa haye aur 2020 taka koshika avasthaon aur phenotype koo paribhaashit karane key liye swarn maanak maanaa jaataa haye. haalaanki pratyek koshika dwara vyakt pratyek aar. aine. aeye. parr poori jaankaari praapt karna asambhav haye, upalabdh samagri kii chhoti matra key kaaran, jeane abhivyakti pattern koo jeane clustering vishleshanon key madhyam seey pahachaana jaa sakata haye. yeh koshika aabaadi key bheetar durlabh koshika prakaron koo ujagar curr sakata haye joe pahale kabhi naheen dekhe gaye honge. udaaharan key liye, neuroblastoma tumour ootak parr scrana-seik karane vaale vaigyaanikon key ack samooh ney ack durlabh paine-neuroblastoma cancer koshika kii pehchaan kii, joe nae chikitsa drishtikon key liye aakarshak hoe sakati haye.
vidhiyaan.
vartamaan scrana-seik protocol mein ekal koshikaon aur unake aar. aine. aeye. koo alag karna shamil haye, aur phir bulk aar. aine. aeye.-seik key roop mein usee charanon kaa paalan karnaah riverse transcription (aar. tii.), pravardhan, pustakalaya utpaadan aur anukraman. praarambhik vidhiyon ney alag-alag koshikaon koo alag-alag kuon mein alag kiya; adhik haal key tareeke ack microfluidic upkaran mein alag-alag koshikaon koo alag-alag boondon mein samaahit karate hain, jahaan riverse transcription pratikriya hoti haye, joe rna koo cdna mein parivartit karati haye. pratyek boond mein ack dna "barcode" hotaa haye joe ekal koshika seey praapt cdna koo vishisht roop seey label kartaa haye. ack baar riverse transcription puuraa hoe jaane key baad, kaee koshikaon key cdna koo anukraman key liye ack saath milaaya jaa sakata haye, kyonki ack vishesh koshika seey pratilekhon kii pehchaan advitiya barcode dwara kii jaati haye.
scrana-seik key liye chunautiyon mein ack koshika mein mrinaa kii praarambhik saapeksh prachurta koo sanrakshit karna aur durlabh pratilekhon kii pehchaan karna shamil haye. riverse transcription charan mahatvapurna haye kyonki aar. tii. pratikriya kii dakshata yeh nirdhaarit karati haye kii koshika kii aar. aine. aeye. aabaadi kaa kitna vishleshan antatah anukramak dwara kiya jaayega. riverse transcriptase kii prakriya aur upayog kii jaane vaali priming ranneetiyaan poorna-lambai see. dee. aine. aeye. utpaadan aur 3 'yaa 5' jeane key anta kii or pakshapaati pustakalayon kii peedhi koo prabhaavit curr sakati hain.
pravardhan charan mein, yaa tou pee. see. aar. yaa inn vitro transcription (aaee. vee. tii.) kaa upayog vartamaan mein see. dee. aine. aeye. koo pravardhit karane key liye kiya jaataa haye. pee. see. aar.-adharit vidhiyon key laabhon mein seey ack poorna-lambai see. dee. aine. aeye. utpann karane kii kshamata haye. haalaanki, vishesh anukramon parr alag-alag pee. see. aar. dakshata (udaaharan key liye, jee. see. samagri aur snapback sanrachana) bhee teji seey pravardhit kii jaa sakati haye, jisase asman coverage key saath pustakalayon kaa utpaadan hotaa haye. doosari or, aaee. vee. tii. dwara utpann pustakalaya pee. see. aar.-prerit anukram purvagrah seey butch sakate hain, vishisht anukramon koo aksham roop seey pratilekhit kiya jaa sakata haye, iss prakaar anukram drop-out yaa apoorna anukram utpann hoe sakate hain.
kaee scrana-seik protocol prakaashit kiye gaye hainh
taang aur anya., 
strt, 
smart-seik, sort-seik, 
cell-seik, krodh-seik,
quartz-seik.
, aur see1-केज।. yeye protocol riverse transcription, see. dee. aine. aeye. sanshleshan aur pravardhan key liye ranneetiyon key sandarbh mein bhinn hotey hain, aur anukram-vishisht barcode (i.ee., umis) yaa ekatrit namoonon koo sansaadhit karane kii kshamata.
2017 mein, ack saath ekal-koshika aim. aar. aine. aeye. aur protein abhivyakti koo maapane key liye dou drishtikon pesh kiye gaye they, jinhen riek-seik aur site-seik key roop mein jaane jaane vaale oligonucleotide-label antibody key madhyam seey maapane key liye. patch-clamp kaa upayog karke electrophysiological recording key baad cellular samagri ekatra karane seey patch-seik vidhi key vikas kii bhee anumati millie haye, joe tantrika vigyaan mein lagaatar aadhar praapt curr rahee haye.
ack boond adharit munch kaa udaaharan-10x vidhi.
ekal koshika aar. aine. aeye. anukraman kaa yeh munch ekal koshikaon koo pakadane aur agali peedhi key anukraman (aine. jee. aiss.) see. dee. aine. aeye. pustakalayon koo taiyar karane key liye microfluidic vibhajan key upayog dwara koshika-darr-koshika aadhar parr pratilekhon kaa vishleshan karane kii anumati detaa haye. boond adharit munch tail kii boond mein ekal koshika koo pakadkar badee sankhya mein vyaktigat koshikaon mein aim. aar. aine. aeye. key badey paimaane parr samanantar anukraman koo saksham banaata haye.
kull milakar, pahale charan mein alag-alag koshikaon koo alag seey pakada jaataa haye aur unhein alag kiya jaataa haye, phir aim. aar. aine. aeye. kaa riverse transcription (aar. tii.) kiya jaataa haye aur see. dee. aine. aeye. library praapt kii jaati haye. aim. aar. aine. aeye. kaa chayan karane key liye, aar. tii. koo deoxythymine (oligo dee. tii.) primer key ekal-fanse anukram key saath kiya jaataa haye joe vishesh roop seey aim. aar. aine. aeye. anuon kii polly (aeye) poonchh koo baandhata haye. baad mein, samvardhit see. dee. aine. aeye. pustakalaya kaa upayog anukraman key liye kiya jaataa haye.
tou, vidhi kaa pahala kadam ekal koshika aavaran aur pustakalaya kii taiyari haye. koshikaon koo ack swachaalit key kaaran jail motee-inn-emulsion (ratna) mein samaahit kiya jaataa haye. inn putikaon koo banaane key liye, swachaalit ack sookshm drav chip kaa upayog kartaa haye aur tail key saath sabhi ghatakon koo jodata haye. pratyek karyatmak ratna mein ack ekal koshika, ack ekal jail beed aur aar. tii. abhikarmak hotey hain. jail beed parr, 4 alag-alag bhaagon seey baney olignonucleotide baandhate hainh pee. see. aar. primer (anukraman key liye aavashyak); 10x barcoded oligonucleotide; advitiya aanvik pahchankarta (yumi) anukram; polyedt anukram (joe polly-adenalated aim. aar. aine. aeye. anuon koo pakadane mein saksham banaata haye). 
pratyek ratna pratikriya putika key bheetar, ack ekal koshika koo liz kiya jaataa haye aur iska pratilekhan vipareet hotaa haye. ack hii koshika seey cdna kii pehchaan ack samanya 10x barcode key kaaran kii jaati haye.
isake alawa, oomis kii sankhya jeane abhivyakti star koo vyakt karati haye aur iska vishleshan atyadhik parivartansheel jeane kaa pataa lagaane mein madad kartaa haye. una aankadon kaa upayog aksar yaa tou cellular phenotype vargeekaran yaa naee upa-jansankhya pehchaan key liye kiya jaataa haye.
munch kaa antim charan anukraman haye. utpann pustakalayon kaa upayog seedhe ekal koshika poore pratilekh anukraman yaa lakshya anukraman karyapravah key liye kiya jaa sakata haye. anukraman roshni dye anukraman vidhi kaa upayog karke kiya jaataa haye. yeh anukraman vidhi sanshleshan (aiss. bee. aiss.) siddhant dwara anukraman aur prativarti dye-terminator key upayog parr adharit haye joe pratyek ekal nucleoted kii pehchaan koo saksham banaata haye.
ack chhor parr pratilekh anukramon koo padhane key liye, aur dusare chhor parr barcode aur yumi, yugmit-anta anukraman pathakon kii aavashyakta hoti haye.
boond-adharit munch apane uchch pravaah kshamata key kaaran durlabh koshika prakaron kaa pataa lagaane kii anumati detaa haye. vaastav mein, ack ekal koshika nilamban seey prati namoone 500 seey 10,000 koshikaaen lii jaati hain. protocol aasaani seey kiya jaataa haye aur 65 pratishat taka kii uchch koshika punarpraapti darr kii anumati detaa haye. boond-adharit munch key vaishvik karyapravah mein 8 ghante lagate hain aur isaliye yeh microwell-adharit vidhi (bee. dee. rhapsody) kii tulanaa mein tej haye, jisamen 10 ghante lagate hain.
haalaanki, yeh kuchh seemaon koo prastut kartaa haye kyonki nae namoonon kii aavashyakta aur kewal 10 pratishat aim. aar. aine. aeye. kaa antim pataa lagaana.
droplet-adharit vidhi aur microwell-adharit vidhi key beach pramukh antar koshikaon key vibhajan key liye upayog kii jaane vaali takneek haye.
seemaaen.
adhikansh aar. aine. aeye.-seik vidhiyaan aim. aar. aine. aeye. koo samriddh karane aur prachur matra mein aur suchanaaheen aar. aar. aine. aeye. koo cumm karane key liye polly (aeye) tail capture parr nirbhar karati hain. iss prakaar, way aksar polyadenylite aim. aar. aine. aeye. anuon koo anukramit karane taka seemit hotey hain. haalaanki, haal key adhyayan abb gair-polly (aeye) aar. aine. aeye. key mahatva kii sarahana karna shuru curr rahey hain, jaise kii jeane abhivyakti viniyaman mein long-noncoding aar. aine. aeye. aur microrna. small-seik ack ekal-koshika vidhi haye joe standhari koshikaon mein chhote aar. aine. aeye. (<300 nucleotide) jaise microrna, trana key tukade aur chhote nucleolar aar. aine. aeye. koo pakadati haye. yeh vidhi "oligonucleotide mask" (joe atyadhik prachur matra mein 5.8s rrna anuon key grahan koo rokta haye) aur aakar chayan key sanyojan kaa upayog karati haye taaki badee rna prajaatiyon jaise kii anya atyadhik prachur matra mein rrna anuon koo baahar rakhaa jaa sakey. badey gair-polly (aeye) rna koo lakshit karane key liye, jaise kii lambe gair-coding mra, histone mra, golakar rna, aur vardhak rna, aakar chayan atyadhik prachur matra mein ribosomal rna anuon (18 aur 28 rra) koo cumm karane key liye lagu naheen hotaa haye. ekal-koshika ramda-seik ack aisee vidhi haye joe vishesh roop seey aar. aar. aine. aeye. anu parr priming seey bachane key liye design kiye gaye "itna yaadrichhik naheen" (aine. aiss. aar.) primer kii upasthiti mein yaadrichhik priming (yaadrichhik visthaapan pravardhan) key saath riverse transcription karke ise praapt karati haye. jabaki yeh vidhi anukraman key liye poori lambai key kull aar. aine. aeye. pratilekhon koo safaltapurvak pakadati haye aur uchch samvedansheelta key saath vibhinn prakaar key gair-polly (aeye) aar. aine. aeye. kaa pataa lagaati haye, isaki kuchh seemaaen hain. nsr primer koo vishisht jeev (mouse) mein rrna anukramon key anusaar saavdhaanipoorvak design kiya gayaa thaa, aur anya prajaatiyon key liye nae primer sett koo design karane mein kaafee prayaas karna hoga. haal hii mein, scadash naamak ack chrisp-adharit vidhi (sankaran dwara prachur matra mein anukramon kaa ekal-koshika hraas) ney ekal-koshika kull aar. aine. aeye.-seik pustakalayon seey aar. aar. aine. aeye. anukramon koo cumm karane key liye ack aur drishtikon kaa pradarshan kiya. 
polyadenylite aim. aar. aine. aeye. kii kamee key kaaran bacteria aur anya procariotes vartamaan mein ekal-koshika aar. aine. aeye.-seik key liye anukool naheen hain. iss prakaar, ekal-koshika aar. aine. aeye.-seik vidhiyon kaa vikas joe polly (aeye) tail capture parr nirbhar naheen karate hain, ekal-koshika resolution sookshm jeev adhyayan koo saksham karane mein bhee sahayak hoga. thoke jeevaanu adhyayan aamtaur parr bacteria parr polyadenylated aim. aar. aine. aeye. kii kamee koo door karane key liye samanya aar. aar. aine. aeye. avakshay koo lagu karate hain, lekin ekal-koshika star parr, ack koshika mein paaya jaane vaalaa kull aar. aine. aeye. bahut chhota hotaa haye. polyadenylated aim. aar. aine. aeye. kii kamee aur ekal bacteria koshikaon mein paae jaane vaale kull aar. aine. aeye. kii kamee dou mahatvapurna baadhaaen hain joe bacteria mein scrana-seik kii tainaati koo seemit karati hain. 
anuprayog.
vikasatmak jeev vigyaan, tantrika vigyaan, oncology, pratirakshavigyan, hriday anusandhaan aur sankraamak roga sahit javic vishayon mein scrana-seik kaa vyaapak roop seey upayog kiya jaa rhaa haye.
machine learning vidhiyon kaa upayog karate hue, bulk aar. aine. aeye.-seey. kyu. seey detta kaa upayog aiss. aar. aine. aeye.-seey. kyu. mein sanket/shore anupaat badhaane key liye kiya gayaa haye. vishesh roop seey, vaigyaanikon ney saha-abhivyakti network banaane key liye paine-cancer dataset seey jeane abhivyakti profile kaa upayog kiya haye, aur phir inhen ekal koshika jeane abhivyakti profile parr lagu kiya haye, transcript staron kaa upayog karke vyaktigat koshikaon mein utparivartan kii upasthiti kaa pataa lagaane key liye ack adhik majboot vidhi praapt kii haye.
kuchh scrana-seik vidhiyon koo ekal koshika sookshmajeevon parr bhee lagu kiya gayaa haye. smart-secue2 kaa upayog ekal koshika eukariotic sookshmajeevon kaa vishleshan karane key liye kiya gayaa haye, lekin choonki yeh polly (aeye) tail capture parr nirbhar kartaa haye, isaliye ise procariotic koshikaon mein lagu naheen kiya gayaa haye. ekal maleria parjivi yaa ekal khameer koshikaon koo anukramit karane key liye drop-seik aur drav aaee. aff. see.-see. 1 upkaranon jaise sookshm-drav drishtikon kaa upayog kiya gayaa haye. ekal-koshika khameer adhyayan ney khameer key namak key tanaav key sampark mein aaney seey pahale aur baad mein isogenic khameer koshikaon mein visham tanaav sahishnuta koo chihnit karane kii maang kii. scrana-seik dwara kaee pratilekhan karakon key ekal-koshika vishleshan seey aabaadi mein vishamta kaa pataa chalaa. inn parinamon seey pataa chalata haye kii jansankhya key ack ansh key liye jeevit rahane kii sambhavana badhaane key liye jansankhya key sadasyon key beach viniyaman bhinn hotaa haye.
procariotic prajaati mein pahala ekal-koshika pratilekh vishleshan terminator exonucles enzyme kaa upayog karke aim. aar. aine. aeye. key chuninda roop seey aar. aar. aine. aeye. aur rolling circle amplification (aar. see. aeye.) koo cumm karane key liye kiya gayaa thaa. iss vidhi mein, ekal-fanse hue dna key siron koo ack vritt banaane key liye ack saath jodaa gayaa thaa, aur parinami loop kaa upayog raikhik rna pravardhan key liye ack template key roop mein kiya gayaa thaa. antim utpaad pustakalaya kaa vishleshan sookshm sarani dwara kiya gayaa thaa, jisamen cumm purvagrah aur achchi coverage thee. haalaanki, aar. see. aeye. kaa aar. aine. aeye.-seik key saath pareekshan naheen kiya gayaa haye, joe aamtaur parr agali peedhi key anukraman koo niyojit kartaa haye. bacteria key liye ekal-koshika aar. aine. aeye.-seik sookshm jeevon key adhyayan key liye atyadhik upayogi hoga. yeh paaramparik thoke metatronscryptomics drishtikon mein saamane aaney vaale muddon koo sambodhit karega, jaise kii cumm prachurta mein maujood prajaatiyon koo pakadane mein whiffle rehna, aur koshika aabaadi key beach vishamta koo hull karane mein whiffle rehna.
scrana-seik ney bhroon aur jeevon key vikas mein kaafee antardrishti pradaan kii haye, jisamen krimi canorhabditis elegans aur punaryoji planarian smidita mediterrania aur exolotal ambistoma maxicanum shamil hain. iss tarah seey maanchitrit kiye jaane vaale pahale kasheruki janwar zebrafish aur "zenopus levis" they. pratyek mamle mein bhroon key kaee charanon kaa adhyayan kiya gayaa, jisase vikas kii poori prakriya koo koshika-darr-koshika aadhar parr mapp kiya jaa sakey. vigyaan ney inn pragati koo varsh 2018 kii safalata key roop mein maanyata dee.
choohon key vrishanon kaa ack aanvik koshika atlus, joe ack ekal-koshika sankalp parr bee. dee. 47-sambandhit vrishan chot mein shamil antarnihit tantra aur maargon kii hamari samajh mein naee antardrishti pradaan karate hue, scrana-seik drishtikon kaa upayog karke bee. dee. 47-prerit pree-tuberal testicular vishaaktata koo paribhaashit karane key liye sthaapit kiya gayaa thaa.
vichaar.
ekal koshikaon kaa algaav.
poore genome pravardhan aur anukraman seey pahale vyaktigat koshikaon koo alag karane key kaee tareeke hain. pratideepti-sakriya koshika chhantaai (faces) ack vyaapak roop seey upayog kiya jaane vaalaa drishtikon haye. vyaktigat koshikaon koo sookshm-manankan dwara bhee ekatra kiya jaa sakata haye, udaaharan key liye kramik mandan dwara yaa ekal koshika kii kataai key liye patch pipet yaa nanotube kaa upayog karke. sookshm-parimanikaran key laabh aasaan aur cumm laagat vaale hain, lekin way sookshmadarshi key tahat koshika prakaron kii galat pehchaan key liye shramsadhya aur atisamvedansheel hain. lazer-capture microdisction (ell. see. aim.) kaa upayog ekal koshikaon koo ikattha karane key liye bhee kiya jaa sakata haye. halanki ell. see. aim. ack ootak key bheetar ack namuna koshika key sthaanik sthaan key gyaan koo sanrakshit kartaa haye, lekin padosi koshikaon seey samagri ekatra kiye binaa ack poori ekal koshika koo pakadana mushkil haye. 
ekal koshika algaav key liye uchch-utpaadan vidhiyon mein sookshm fluidics bhee shamil hain. donon pahalu aur sookshm drav sateek, swachaalit aur nishpaksh namoonon koo alag karane mein saksham hain. haalaanki, donon vidhiyon key liye pahale apane sookshm vatavaran seey koshikaon koo alag karane kii aavashyakta hoti haye, jisase aar. aine. aeye. abhivyakti vishleshan mein pratilekhan profile mein gadbadi hoti haye.
vishleshan kiye jaane vaale koshikaon kii sankhya.
scrana-seik.
aam taur parr, ack vishisht bulk cell aar. aine. aeye.-sequencing (aar. aine. aeye.-secue) prayog key liye, duss million reed utpann hotey hain aur 50 reed prati key. bee. reed (aar. pee. key. aim.) kii seema seey adhik vaale jeane koo vyakt maanaa jaataa haye. 1key. bee. lambe jeane key liye, yeh poison vitaran kii dhaarana key tahat 500 reeding aur 4 pratishat kii bhinnata (see. vee.) key nyuntam gunank seey mell khaata haye. 200, 000 aim. aar. aine. aeye. yukt ack vishisht standhari koshika key liye, iss nyuntam see. vee. moolya koo praapt karane key liye cumm seey cumm 50 ekal koshikaon seey anukraman detta koo ekatra karane kii aavashyakta hoti haye. haalaanki, prayogon mein riverse transcription aur anya shore kii dakshata key kaaran, sateek abhivyakti vishleshan aur koshika-prakaar kii pehchaan key liye adhik koshikaon kii aavashyakta hoti haye.

