1 lace Simbuza ibikoresho byubushakashatsi mugihe

Shiraho (NTC) kugenzura nabi hanyuma ubisubiremo inshuro nyinshi.Bimaze kugaragara ko muri PCR hari ibicuruzwa byanduye muri laboratoire, simbuza ibikoresho byose byubushakashatsi mugihe.Nka: ongera ushushe kandi utegure primers, ongera uhindure umuyoboro wa pipette, umuyoboro wa EP, ddH2O, nibindi, usimbuze umuyoboro mushya, hanyuma utiza izindi laboratoire gukora ubushakashatsi bwa PCR.Laboratoire yanduye igomba guhumeka no gushyirwamo imirasire ya ultraviolet kugeza igihe PCR yanduye ikuweho mbere yo gukomeza ubushakashatsi.

2 tend Ongera igihe cyo kwerekana UV

Birakwiye ko tumenya ko kugirango ukureho ADN yanduye, imirasire ya ultraviolet isanzwe igomba kongerwa amasaha 2 kurenza uko bisanzwe.Nubwo bimeze bityo, ingaruka za UV irrasiyo mugukuraho uduce duto (munsi ya 200bp) ya ADN yanduye ntago ari nziza.

Uburebure bwa Ultraviolet (nm) muri rusange ni 254 / 300nm, kandi birahagije kurasa muminota 30.Twabibutsa ko mugihe uhisemo UV kugirango ikureho kwanduza ibicuruzwa bisigaye bya PCR, uburebure bwibicuruzwa bya PCR hamwe nogukwirakwiza ibishingwe mubicuruzwa bikurikirana.Imirasire ya UV ikora gusa kubice birebire hejuru ya 500 bp, kandi ntigira ingaruka nke kubice bigufi.

Mugihe cya UV irrasiyoya, pyrimidine ishingiro mubicuruzwa bya PCR bizakora ibipimo.Ibipimo birashobora guhagarika kwaguka, ariko ntabwo pyrimidine zose ziri mumurongo wa ADN zishobora gukora ibipimo, kandi irrasiyo ya UV nayo irashobora kumena ibipimo..Urwego rwo gushiraho dimer rushingiye kuburebure bwa UV, ubwoko bwa pyrimidine dimer hamwe nurutonde rwa nucleotide yegeranye na dimer.Kubwibyo, niba PCR yongerewe ibice ari bito, birasabwa gukoresha sisitemu yo kwanduza ibicuruzwa UNG irwanya PCR.

3 ： Ikoreshwa rya ADN zikoreshwa cyane

Aerosole ikorwa byoroshye mugihe imiyoboro yongeweho, bigoye kuyirinda, kandi izatuza vuba.Kubwibyo, nta gushidikanya ko ari amahitamo meza yo gukoresha kenshi ibyuma byangiza umwanda wa ADN kugirango wirinde ikwirakwizwa rya ADN.

4 ： Koresha uburyo bwa UNG bwo kurwanya umwanda

Nyuma yo kwanduza ibicuruzwa bya PCR bimaze gukurwaho, laboratoire yipimisha irashobora gukoresha sisitemu yo kwanduza UNG anti-PCR kugirango ikumire ibicuruzwa bya PCR.Muri laboratoire rusange, urashobora gukora ibice byoroshye byubushakashatsi, gutandukanya byimazeyo ibicuruzwa bya PCR nibindi bice, ugashyiraho amategeko n'amabwiriza ya laboratoire, kandi ugakora amahugurwa ajyanye no gukumira ko PCR yanduye.

Ibyifuzo: Gushiraho laboratoire ya PCR ishyize mu gaciro, kubungabunga ibidukikije byiza bya PCR, gushyiraho uburyo busanzwe bwo gukora PCR, no gutsimbataza ubumenyi bukomeye bwibikorwa byabashakashatsi ni urufunguzo rwo gukumira cyangwa kugabanya umwanda w’ubushakashatsi bwa PCR.