
recombinage polymerase amplification

recombinase polymerase amplification (arpa) anedi polymerase chain reaction (pcr) kii isothermal pratyamnaayamgaa oke gottam. arpa praticharyaku riverse transcriptage enjaimnu jodinchadam dwara idhi cdnnu utpatti cheyadaaniki pratyeka dasha avasaram lekunda rnatho paatu dnnu gurtinchagaladu. idhi isothermal kabatti, arpa pcr kante chaalaa saralamaina parikaraalanu upayoginchagaladu, dheeniki thermal cycler avasaram. 37-42 °C ushnogratala vadda uttamamgaa panicheestuu, inca panicheestuu, marinta nemmadigaa unnappatikee, gadhi ushnograta vadda arpa praticharyalanu siddhaantaparamgaa kevalam gottanni pattukovadam dwara twaragaa amalu cheyavachu. idhi takkuva kharchutho kuudina, vegavantamaina, point-off-cair molecular pareekshalanu abhivruddhi cheyadaaniki aarpienu adbhutamaina abhyardhigaa chestundi. konni pii. sii. aarr pareekshalu mariyu aarr. tii-lamp pareeksha dwara tappipoyina takkuva saandrata namuunaalanu gurtinchadam dwara, rift valley feavour virus yokka molecular detection yokka antarjaateeya naanhyata anchanaa alaage uttama aarr. tii-pii. sii. aarr pareekshalu nirvahinchabadataayi.
aarpienu twistdx limited abhivruddhi chesi praarambhinchindi. (gatamlo asm scientific limited anie pilicheevaaru), idhi britanloni cambridgelo unna oka biotechnology samstha.
saanketikata.
arpa prakriya moodu pradhana enjaimlanu upayogistundi-oka recombinase, oka single-stranded dna-binding protein (ssb) mariyu strand-displacing polymerase.
recombinagelu duplexes dnalo oligonucleotide primerlanu homologus sequensto jatha cheyagalavu.
ssb dna yokka sthaanabhramsham chendina tantuvulatoe bandhistundi mariyu primerlu sthaanabhramsham chendakunda nirodhistundi.
chivaraku, strand sthaanabhramsham polymerase anedi dna samshlaeshanhanu praarambhistundi, ikkada primer lakshya dnaku kattubadi untundi.
rendu pratyarthi primerlanu upayoginchadam dwara, chaalaa varaku pcr laaga, lakshya kramam nijamgaa unte, ghaataanka dna amplification praticharya praarambhinchabadutundi. vistarananu praarambhinchadaaniki ushna ledhaa rasayana draveebhavanam vanti itara namuunaa taarumaaru avasaram ledhu. saraina ushnogratala vadda (37-42 °C), praticharya vaegamgaa munduku saagutundi mariyu viral janyu DNA ledhaa RNA, vyadhikaaraka bacteria janyu DNA, alaage swalpakaalika optamer DNA nuu vaegamgaa gurtinchadaaniki, saadhaaranamgaa 10 nimishaallopu, gurtinchadagina sthaayilaku konni lakshya kaapeela nundi nirdishta DNA vistaranaku daariteestundi.
adanapu kaaryaacharananu andinchadaaniki moodu pradhana arpa enjaimlanu marinta enjaimlatho bharti cheyavachu. exonucles III nii jodinchadam anedi real-time, fluorescence detection kosam real-time pcr maadirigaa exo probnu upayoginchadaaniki anumatistundi. endonuclease IV nii jodinchadam antey vijayavantamaina vistarana yokka parshwa pravaaha patteeni gurtinchadaaniki nfo probnu upayoginchavacchu. 37-42 °C vadda panichese riverse transcriptagenu jodinchinatlayithe, rnanii riverse transcript cheyavachu mariyu utpatti cheyabadina cdna annintinee oke dasalo vistarinchavacchu. prastutam riverse transcriptesto kuudina arpa yokka twistamp exo version matrame andubaatulo undhi, ayithe adhe prabhavanni utpatti cheyadaaniki viniyogadaarulu itara twistamp praticharyalanu riverse transcriptesto bharti cheyavachu.
pcr maadirigaane, anni rakaala arpa praticharyalanu marinta primer/probe jatalanu jodinchadam dwara bahulamgaa cheyavachu, idhi oke gottamlo bahula vislaeshanhalanu gurtinchadaaniki ledhaa antargata niyantrananu anumatistundi.
itara vistarana paddhathulatho sambandham.
paramaanu roganirdhaarana saanketikatagaa abhivruddhi cheyabadina aneka isothermal nucleic assid amplification techniclalo arpa okati, tarachugaa pcrku sambandhinchi avasaramaina prayogasaala parikaraalanu saralikrutam chese lakshyamtho. itara isothermal amplification technicla pakshika jaabitaalo lamp, nasba, helicase-dependent amplification (hcda) mariyu nicking enjaim amplification reaction (sameepamlo) unnaayi. primer design mariyu reaction mechanism yokka pratyekatalalo paddhatulu bhinnamgaa untaayi, mariyu konni sandarbhaallo (arpa vantivi) rendu ledhaa anthakante ekkuva enjaimla cactailsnu upayogistaayi. arpa maadirigaane, eee paddhatulu chaalaa varaku saralikruta parikaraala saamardhyamtho vegavantamaina vistarana samayaalanu andistaayi mariyu pcrnu nirodhinche suddhi cheyani namoonaalalo padaarthaalaku nirodhakatanu nivedistaayi. vistarana samayaniki sambandhinchi, vegavantamaina ushnograta ryaamplatoe kuudina aadhunika thermocyclerlu pcr vistarana samayanni 30 nimishala kante takkuvagaa tagginchagalavu, mukhyamgaa saampradaaya moodu-ushnograta protokaallaku badulugaa dwandwa-ushnograta saiklingnu upayoginche chinna amplicanla kosam. adanamgaa, namuunaa tayari yokka demandlanu (avasaramaite, lysis mariyu dna ledhaa rna velikiteethatho sahaa) saanketikataku antarleenamgaa unna mottam samayam mariyu sanklishtatalo bhaagamgaa pariganinchaali. eee avasaralu saanketikatato paatu nirdishta lakshyam mariyu namuunaa rakanni batti maarutuu untaayi.
pcr thoo poliste, primer mariyu probe design kosam maargadarsakaalu arpa kosam takkuva sthirapadathaayi, mariyu kontha trial und error teesukovachhu, ayithe iteevali phalitaalu praamaanika pcr primerlu kooda pani cheyagalavani suuchistunnaayi. (aichhika) antargata fluorogenic probto munduku mariyu venukaku praimartho sarihaddulugaa unna vivikta amplican yokka saadhaarana suutram pcrnii pooli untundi. pcr primerlanu neerugaa arplo upayoginchavacchu, conny vaati takkuva podavu antey punarekeekarana raetlu takkuvagaa untaayi mariyu arpa mukhyamgaa sunnitamainadi ledhaa vegavantamainadi kaadu. samarthavanthamaina recombinase filament nirmaanam mariyu arpa paniteeru kosam saadhaaranamgaa 30-38 bays primerlu avasaram. idhi adanapu design parimitulaku lobadi pedda sankhyalo primerlanu upayoginche deepam vanti konni itara paddhatulaku viruddhamgaa untundi. arpa yokka asalu 2006 nivedika kriyaatmaka praticharya bhagala samitini vivarinchinappatikii, twistamp kitt yokka prastuta (yajamanya) suutreekarana "gananeeyamgaa bhinnamgaa" untundi mariyu twistdx sarafaraadaaru nundi matrame andubaatulo untundi. idhi chaalaa mandhi sarafaraadaarula nundi labhinche pii. sii. aarr. kosam praticharya misramaalato polchadam, ledhaa praticharya misramam yokka kuurpu swechchagaa prachurinchabade deepam ledhaa nasba, parisoedhakulu chavakaina padaardhaala nundi vaari swantha anukuuleekarinchina "kitlu" srushtinchadaaniki veelu kalpistundi.
prachurinchabadina shaastreeya saahityamlo saadhaaranamgaa arpa, hcda mariyu deepam vanti isothermal amplification technicla paniteerunu okadaanikokati sambandhinchi vivaranaatmaka poolika undadu, tarachugaa oke isothermal tekniknu "goaled standard" pcr aselo poolchutaaru. idhi tayaareedaarulu, aavishkartalu ledhaa pratipaadakula vaadanala nundi swatantramgaa eee paddhatula yokka yogyatalanu anchanaa veyadam kashtataram chestundi. antekakunda, edainaa amplification technic yokka paniteeru lakshanaalanu primer design nundi vidadeeyadam kashtam arpa kosam oka lakshyam kosam sett chesina "manchi" primer adhe lakshyam kosam sett chesina "paelavamaina" lamp primer kante vaegamgaa amplification ledhaa marinta sunnitamaina gurtimpunu ivvavacchu, conny vaerae lakshyam kosam ververu primer setlaku dheeniki viruddhamgaa undavacchu. oka minahaayimpu emitante, schmallenberg virus mariyu bovin viral diarrhoea vairasnu gurtinchadaaniki arty-qpcr, arty-lamp mariyu aarpienu polchina iteevali adhyayanam, idhi prati amplification teknikku balaalu mariyu balaheenatalu unnayani, idhi lakshyanni batti maaravacchu, mariyu andubaatulo unna amplification technicla lakshanaalanu prati application yokka avasaraalatoe kalipi anchanaa veyaalsina avasaram undani samarthavantamgaa teliyajestundi. pcr mariyu itara amplification technic maadirigaa, paelavamaina paniteeru kaligina primer setlu nivedinchadaaniki arhamainaviga arudugaa pariganinchabadatam valla, prachurana pakshapaatam spashtamgaa undhi.

