Ọtụtụ enzymes a na-ejikarị na nnwale PCR

Mmeghachi omume yinye nke polymerase, nke akpọchiri dị kaPCRn'asụsụ bekee, bụ usoro ihe ọmụmụ ihe omimi nke a na-eji amụba iberibe DNA a kapịrị ọnụ.Enwere ike were ya dị ka mkpụrụ ndụ DNA pụrụ iche n'èzí ahụ, nke nwere ike ịbawanye ntakịrị DNA.N'oge dumPCRUsoro mmeghachi omume, otu klas nke ihe na-arụ ọrụ dị mkpa - enzymes.

1. Nke DNA

Na nnwale na mmalite ụbọchị nkePCRNdị ọkà mmụta sayensị na-eji Escherichia coli DNA polymerase I, Ma enwere nsogbu na enzyme a: ọ dị mkpa iji mejupụta enzyme ọhụrụ oge ọ bụla a na-eme okirikiri, nke na-eme ka ọrụ ahụ dị ntakịrị mgbagwoju anya na ọ na-esiri ike ịmalite ngwa ngwa ngwa ngwa.A doziri nsogbu a mgbe ndị ọkà mmụta sayensị kewapụrụ Taq DNA polymerase na mberede na Thermus aquaticus na 1988. Kemgbe ahụ, mmụba akpaka nke DNA aghọwo eziokwu.Nchọpụta nke enzyme a na-emekwaPCRtechnology a adaba, bara uru na eluigwe na ala technology.Ugbu a, Taq DNA polymerase bụ polymerase kachasị na ngwa DNA.

2. PfuDNA

Dịka ekwuru n'elu, Taq DNase nwere nnukwu ahụhụ, ya mere ndị ọkà mmụta sayensị agbanweela Taq DNA polymerase ruo n'ókè ụfọdụ iji zere mmụba na-abụghị nke akọwapụtara n'ihi enweghị nkwekọrịta, na-ebute nsonaazụ ule na-ezighi ezi.Mana ngbanwe nke Taq DNA polymerase nwere ike igbochi ọrụ DNA polymerase na okpomọkụ ụlọ.PfuDNA polymerase nwere ike mejupụta ihe ọghọm dị n'elu nke Taq DNA polymerase, ka enwere ike ịme mmeghachi omume PCR nke ọma, yana enwere ike ịkwalite ọnụego ihe ịga nke ọma nke nkwalite mkpụrụ ndụ ihe nketa.

3. Reverse Transcriptase

Achọpụtara reverse transcriptase na 1970. Enzyme a na-eji RNA dị ka ndebiri, dNTP dị ka mkpụrụ, na-agbaso ụkpụrụ nke njikọta isi, ma na-emepụta otu eriri DNA na-ejikọta na template RNA na ntụziaka 5'-3′.Reverse transcriptase na-adabere na ọrụ DNA polymerase sitere na DNA ma ọ bụ ndebiri RNA ya mere enweghị ọrụ 3′-5′ exonuclease.Agbanyeghị, ọ nwere ọrụ RNase H, nke na-amachi ogologo njikọ nke reverse transcriptase ruo n'ókè ụfọdụ.N'ihi obere iguzosi ike n'ezi ihe na thermostability nke ọhịa reverse transcriptase, ndị ọkà mmụta sayensị mezikwara ya.

MakaPCRnnwale, ndị bụ isi consumables bụ: onye PCR tube, 4/8-strip PCR tube, PCR efere.

nke LabioPCR ihe eji eme ihenwere ihe ndị auru:

PCR efere: Ndakọrịta cycler sara mbara;Ihe dị elu, njirimara dị mfe;nke ọma fluorescence echiche; dị mmannyefe okpomọkụ;DNase enwetara ikike, RNase, DNA, ihe mgbochi PCR, yana pyrogen n'efu.

Igwe PCR nke ọ bụla: Evaporation na-eguzogide; dị mmannyefe okpomọkụ;ọmarịcha anya anya;DNase enwetara, RNase, DNA, PCR inhibitors, na pyrogen anwalela.

4/8-eriri PCR tubes: Mgbidi dị oke mkpa; nkọwa dị elu; ezigbo ihe nlegharị anya nke fluorescence;nwere ike iji ya na ụlọ ọrụ ọgwụ, ụlọ ọrụ nri, na usoro ihe ọmụmụ ihe omimi; ihe dị elu, ihe na-amaghị nwoke PP; DNase enwetara ikike, RNase, DNA, PCR inhibitors, na pyrogen anwalela.