Imashini ya PCR | Urumva rwose?

Ikoranabuhanga rya PCR ryegukanye igihembo cyitiriwe Nobel

Mu 1993, umuhanga w’umunyamerika Mulis yahawe igihembo cyitiriwe Nobel muri Chimie, kandi ibyo yagezeho ni uguhimba ikoranabuhanga rya PCR.Ubumaji bwa tekinoroji ya PCR buri mubiranga bikurikira: Icya mbere, ingano ya ADN igomba kongerwa ni nto cyane, kandi mubyukuri ko molekile imwe ishobora gukoreshwa mugukomeza;icya kabiri, amplification imikorere ni myinshi, kandi ingano ya gen igenewe iragaragara.Amplification, inshuro zirenga miliyoni 10 mumasaha make.Ubu igikoresho cya PCR cyakoreshejwe cyane mubushakashatsi bwa siyanse yubuzima nibindi byinshi.

Moderi zitandukanye hamwe nabakora ibicuruzwa byamagare yumuriro barashobora kwerekana imikorere itandukanye no gusubiramo.Itandukaniro ntirigira ingaruka kuri PCR gusa ahubwo rinagira ingaruka no kumenya amakuru yabonetse.Gusobanukirwa ibiranga imashini ya PCR birashobora kudufasha kwerekana intsinzi yubushakashatsi bwacu.

Ubushuhe

Ubushuhe bwubushuhe bwumuriro wumuriro wumuriro burashobora gufata umwanzuro kugirango PCR igerweho cyangwa itsinzwe.Ubushyuhe-bwiza-burigihe ubushyuhe bwo guhagarika ubushyuhe nabwo ni ngombwa kugirango tubone ibisubizo byizewe kandi byororoka PCR.

Bumwe mu buryo bwo kwemeza neza ubushyuhe ni ugupima kenshi ukoresheje ibikoresho byo kugenzura ubushyuhe no kongera gusuzuma nkuko bikenewe numuhanga wabihuguriwe.Ibizamini byo kugenzura ubushyuhe bikoreshwa muburyo bukurikira:

Nibyiza-neza neza ugereranije nubushyuhe muburyo bwa isothermal

Nibyiza-neza neza ugereranije nubushyuhe nyuma yubushyuhe

Shyushya Ubushyuhe Bwuzuye Ubushyuhe

Kugenzura Ubushyuhe Bwambere

Kugenzura ubushyuhe bwa Gradient nigikorwa cyibikoresho bya PCR byorohereza optimizme ya primer annealing muri PCR.Intego ya gradient igenamigambi ni ukugera ku bushyuhe butandukanye hagati ya module, kandi na ≥2 ° C ubushyuhe bwiyongera kandi bukagwa hagati ya buri nkingi, ubushyuhe butandukanye burashobora kugeragezwa icyarimwe kugirango ubone ubushyuhe bwiza bwa primer annealing.Mubyukuri, igipimo cyukuri kigera ku bushyuhe bwumurongo hagati ya module.

Nyamara, ibisanzwe bisanzwe bigenda byerekana ubushyuhe bwumuriro bikoresha ubushyuhe bumwe hamwe no kugenzura ubushyuhe binyuze mubintu bibiri byo gushyushya no gukonjesha biri kumpande zombi, akenshi bikavamo imbogamizi zikurikira:

Ubushyuhe bubiri gusa bushobora gushyirwaho: ubushyuhe bwo hejuru nubushyuhe bwo hasi bwa primer annealing bishyirwa kumpera zombi za module yubushyuhe, kandi imiterere yubushyuhe ntishobora kugerwaho hagati ya modul

Bitewe no guhana ubushyuhe hagati yinkingi zitandukanye, ubushyuhe buri hagati yuturere dutandukanye kuri module birashoboka cyane gukurikira sigmoidal curve aho kuba umurongo nyawo.

Ubushyuhe bw'icyitegererezo

Ubushobozi bwumukino wumukino wo kugenzura ubushyuhe bwicyitegererezo ni ngombwa cyane kubisubizo bya PCR.Ibikoresho byihariye nkibipimo byihuta, gufata igihe, na algorithms ningirakamaro muguhishurira ubushyuhe bwikitegererezo.

Igipimo cyo gushyushya no gukonjesha imashini ya PCR bivuze ko ubushyuhe buhinduka hagati yintambwe za PCR zibaho mugihe runaka.Kubera ko bifata igihe runaka kugirango ubushyuhe bwimurwe kuva muri module kuri sample, igipimo nyacyo cyo gushyushya no gukonjesha icyitegererezo kizatinda.Kubwibyo, ibisobanuro byihuta ryubushyuhe bigomba gutandukanywa no gusobanuka.

Igipimo ntarengwa cyangwa impinga yerekana igipimo cyerekana ubushyuhe bwihuse module ishobora kugeraho mugihe gito cyane mugihe cyo gutambuka.

Ikigereranyo cyo guhagarika igipimo cyerekana igipimo cyimihindagurikire yubushyuhe mugihe kirekire kandi bizatanga igipimo cyerekana umuvuduko wimashini ya PCR.

Igipimo ntarengwa cyo gushyushya no gukonjesha hamwe nimpuzandengo yicyitegererezo cyo gushyushya no gukonjesha byerekana ubushyuhe nyabwo bwabonetse nicyitegererezo.Igipimo cyo gushyushya no gukonjesha bizatanga igereranya ryukuri ryimikorere yimashini ya PCR ningaruka zishobora kugira kubisubizo bya PCR.

Mugihe ukora umusimbuzi wamagare, birasabwa gukoresha igikoresho gifite igipimo cyikigereranyo cyerekana uburyo bwabanje kugirango bisimburwe byoroshye ningaruka ntoya kuri PCR isubirwamo.

Amagare yubushyuhe agomba gutegurwa mugihe cyintambwe gusa nyuma yicyitegererezo kigeze ku bushyuhe bwagenwe.Muri ubu buryo, igihe icyitegererezo cyagumishijwe ku bushyuhe bwagenwe bizakomeza kubungabungwa neza hamwe nuburyo bukurikiranye busabwa muburyo bwo gukora.

Abatwara amagare yubushyuhe bakunze gukoresha imibare igoye ya algorithms kugirango barebe ko ingero zishobora kugera ku bushyuhe bwihuse ukurikije gahunda yateguwe.Ukurikije ingano ya sisitemu ya reaction hamwe nubunini bwa plastiki ya PCR yakoreshejwe, algorithm irashobora guhanura ubushyuhe bwikitegererezo nigihe bizatwara kugirango ubushyuhe bwashyizweho.Wishingikirije kuri algorithms, mugihe cyo gushyushya cyangwa gukonjesha cyumuriro wumuriro wumuriro, ubushyuhe bwo guhagarika ubusanzwe burenga agaciro kagenwe binyuze munzira yiswe amashanyarazi yumuriro cyangwa munsi.Igenamiterere nk'iryo ryemeza ko icyitegererezo kigera ku bushyuhe bwashyizweho vuba bishoboka nta kurasa cyangwa kurasa ubwabyo.

Ubushakashatsi bwinjira

Ibintu bishobora kongera ibicuruzwa byumukino wamagare harimo ibipimo byihuta, ibishushanyo mbonera byahagaritswe, hamwe no guhuza ibyuma byikora.

Igipimo cyo gushyushya no gukonjesha cyicyuma cyumuriro cyerekana umuvuduko ugera kubushyuhe bwagenwe.Nibyihuta ubushyuhe buzamuka kandi bugabanuka, PCR izagenda yihuta, bivuze ko ubushakashatsi bwinshi bushobora kurangira mugihe runaka.Byongeye kandi, ubushakashatsi bushobora kwihuta ukoresheje polymerase ya ADN yihuse.

Igishushanyo mbonera cyumuriro wumuriro nacyo ningirakamaro kubushakashatsi bwa PCR.Kurugero, module isimburwa yemerera guhinduka mumibare yintangarugero kuri buri kwiruka.Mubyongeyeho, gushyushya module hamwe numuntu kugenzurwa kugiti cye nibyiza mugukoresha progaramu zitandukanye za PCR icyarimwe kuri cycler imwe yumuriro.

Kubikoresho byikora cyane-byinjira PCR, software igenzura sisitemu yo gutunganya imiyoboro igomba kuba programme kandi igahuzwa.Sisitemu yikora ni nziza yo gukora ibintu byinshi-byinjira muri PCR kuko birashobora gukoreshwa ubudahwema no gutabara kwabantu, bityo bikagabanya igihe gikenewe cyo gushiraho intoki no kongera umubare wibisubizo mugihe runaka.

Kwizerwa, Kuramba hamwe nubwishingizi bufite ireme bwamagare yubushyuhe

Usibye imikorere nubushobozi bwo kwinjiza, imashini ya PCR igomba kandi kuba ishobora kwihanganira imikoreshereze imwe n'imwe, guhangayikishwa n’ibidukikije, hamwe nuburyo bwo kohereza.Bamwe mubakora ibicuruzwa bashobora gutanga raporo yukuntu igikoresho gikora ibizamini byo kwizerwa no kuramba.Ibikoresho bya PCR bihuye bikubiyemo:

Kwizerwa: Ibikoresho bya mashini bikoreshwa mugukora ibizamini inshuro nyinshi kubikoresho bikoreshwa kenshi nk'ibipfundikizo by'ubushyuhe, imbaho ​​zo kugenzura / gukoraho, hamwe n'ubushyuhe bwo gusiganwa ku magare.

Umuvuduko w’ibidukikije: Ibyumba by’ibidukikije birashobora gukoreshwa mu kwigana imiterere itandukanye y’ubushakashatsi busanzwe, nkubushyuhe, ubushuhe.

Ikizamini cyo kohereza: Ikizamini gikomeye cyo guhinda umushyitsi no kunyeganyega birashobora gukorwa hakurikijwe amahame y’ishyirahamwe mpuzamahanga ryo gutwara ibicuruzwa kugira ngo igikoresho gishobore kugera mu mikorere yangiritse.

Garanti na serivisi yo kubungabunga imashini ya PCR

Nubwo kwizerwa gukomeye no kugeragezwa kuramba, abanyamagare yumuriro byanze bikunze bafite ibibazo bya tekiniki mubuzima bwigikoresho.Kubwamahoro yo mumutima, garanti yuwabikoze, serivisi, no kuyitaho bigomba kwitabwaho mugihe uguze igikoresho.

Ihinduka rya serivisi nko kurubuga / gusubira mu ruganda, serivisi za kure zo kugenzura, hamwe nibikoresho bisimburwa mugikorwa cyo kubungabunga, nibindi, kugirango bigabanye ingaruka kumikorere.

Uburebure bwigihe cya garanti, igihe cyo guhinduka kwa serivisi, kugerwaho ninkunga ya tekiniki, hamwe nubuhanga bwabakozi bunganira umwuga.

Ibishoboka byo gushiraho ibikoresho, imikorere, ubufatanye, no kugenzura kugirango byuzuze laboratoire hamwe nibisabwa bijyanye n'amategeko.Serivise zo gufata neza nko kugenzura ubushyuhe, kugerageza, na kalibrasi zirahari kugirango tumenye neza ko igikoresho gikora neza hamwe nibipimo bihuye.

Ibicuruzwa bifitanye isano: