PCR, kapa polymerase chain reaction, ke mokhoa o sebelisoang ho holisa tatelano ea DNA.E ile ea qalisoa ka lekhetlo la pele lilemong tsa bo-1980 ke Kary Mullis, ea ileng a fuoa Khau ea Nobel ea Chemistry ka 1993 bakeng sa mosebetsi oa hae.PCR e fetotse baeloji ea limolek'hule, e nolofalletsa bafuputsi ho holisa DNA ho tsoa lisampoleng tse nyane le ho e ithuta ka botlalo.
PCR ke ts'ebetso ea mehato e meraro e etsahalang ho sebaesekele sa mocheso, mochini o ka fetolang mocheso oa motsoako oa karabelo ka potlako.Mehato e meraro ke denaturation, annealing, le extension.
Mohato oa pele, denaturation, DNA e nang le mahlakore a mabeli e futhumetse mocheso o phahameng (hangata o ka bang 95 ° C) ho senya li-bond tsa hydrogen tse tšoarang likhoele tse peli hammoho.Sena se etsa hore ho be le limolek'hule tse peli tsa DNA tse mothapo o le mong.
Mohatong oa bobeli, annealing, mocheso o theoleloa ho pota 55 ° C ho lumella li-primers ho ikamahanya le tatellano e tlatsetsang ho DNA ea mokha o le mong.Li-primers ke likotoana tse khutšoane tsa DNA tse etselitsoeng ho tsamaisana le tatellano ea thahasello ho DNA e lebisitsoeng.
Mohato oa boraro, katoloso, mocheso o phahamisetsoa ho pota 72 ° C ho lumella Taq polymerase (mofuta oa DNA polymerase) ho kopanya tlhale e ncha ea DNA ho tloha ho li-primers.Taq polymerase e tsoa ho baktheria e phelang lilibeng tse chesang 'me e khona ho mamella mocheso o phahameng o sebelisoang ho PCR.
Kamora potoloho e le 'ngoe ea PCR, sephetho ke likopi tse peli tsa tatellano ea DNA e reriloeng.Ka ho pheta-pheta mehato e meraro bakeng sa lipotoloho tse 'maloa (ka tloaelo 30-40), palo ea likopi tsa tatellano ea DNA e lebisitsoeng e ka eketsoa ka sekhahla.Sena se bolela hore esita le karolo e nyenyane ea DNA e qalang e ka holisoa hore e hlahise likopi tse limilione kapa tse limilione tse likete.
PCR e na le lits'ebetso tse ngata tsa lipatlisiso le tlhahlobo ea mafu.E sebelisoa ho liphatsa tsa lefutso ho ithuta tšebetso ea liphatsa tsa lefutso le liphetoho, ho tsa forensics ho sekaseka bopaki ba DNA, tlhahlobo ea mafu a tšoaetsanoang ho bona boteng ba likokoana-hloko, le tlhahlobong ea bakhachane ho hlahloba mathata a lefutso maseeng.
PCR e boetse e hlophisitsoe hore e sebelisoe mefuteng e mengata, joalo ka quantitative PCR (qPCR), e lumellang hore palo ea DNA e lekanngoe le ho khutlisa transcription PCR (RT-PCR), e ka sebelisoang ho holisa tatelano ea RNA.
Leha e na le lits'ebetso tse ngata, PCR e na le meeli.E hloka tsebo ea tatellano ea sepheo le moralo oa li-primers tse nepahetseng, 'me e ka ba le phoso haeba maemo a karabelo a sa ntlafatsoe ka nepo.Leha ho le joalo, ka moralo le ts'ebetso e hlokolosi ea liteko, PCR e ntse e le e 'ngoe ea lisebelisoa tse matla ka ho fetisisa ho baeloji ea limolek'hule.
Nako ea poso: Feb-22-2023