Ama-enzyme amaningana avame ukusetshenziswa ekuhlolweni kwe-PCR

I-Polymerase chain reaction, efushanisiwe ngokuthiI-PCRngesiNgisi, indlela yebhayoloji yamangqamuzana esetshenziselwa ukukhulisa izingcezu ezithile ze-DNA.Kungabhekwa njengokuphindaphinda kwe-DNA okukhethekile ngaphandle komzimba, okungakhulisa kakhulu inani elincane kakhulu le-DNA.Ngesikhathi sonkeI-PCRinqubo yokusabela, isigaba esisodwa sezinto sidlala indima ebalulekile - ama-enzyme.

1. Taq DNA

Ezivivinyweni ezinsukwini zokuqala zeI-PCR, ososayensi basebenzise i-Escherichia coli DNA polymerase I, Kodwa kunenkinga ngale enzyme: idinga ukugcwalisa i-enzyme entsha njalo lapho umjikelezo wenziwa, okwenza izinyathelo zokusebenza zibe nzima kancane futhi kunzima ukuzikhulisa ngokugcwele ngokuzenzakalelayo.Le nkinga yaxazululwa ngemva kokuba ososayensi behlukanise ngephutha i-Taq DNA polymerase ku-Thermus aquaticus ngo-1988. Kusukela lapho, ukukhuliswa okuzenzakalelayo kwe-DNA kuye kwaba ngokoqobo.Ukutholakala kwale enzyme nakho kwenzaI-PCRubuchwepheshe ubuchwepheshe obulula, obusebenzayo kanye nobomhlaba wonke.Njengamanje, i-Taq DNA polymerase iyi-polymerase evame kakhulu kumakhithi e-DNA.

2. PfuDNA

Njengoba kushiwo ngenhla, i-Taq DNase inesiphazamisi esikhulu, ngakho ososayensi baye bashintsha i-Taq DNA polymerase ngezinga elithile ukuze bagweme ukukhulisa okungaqondile ngenxa yokungafani, okuholela emiphumeleni yokuhlola engalungile.Kodwa ukuguqulwa kwe-Taq DNA polymerase kungavimbela umsebenzi we-DNA polymerase ekamelweni lokushisa.I-PfuDNA polymerase ingakwazi ukwenza kahle lokhu okungalungile okungenhla kwe-Taq DNA polymerase, ukuze ukusabela kwe-PCR kwenziwe ngokujwayelekile, kanye nezinga lempumelelo lokukhulisa isakhi sofuzo okuhlosiwe lingathuthukiswa ngempumelelo.

3. I-Reverse Transcriptase

I-Reverse transcriptase yatholwa ngo-1970. Le enzyme isebenzisa i-RNA njengesifanekiso, i-dNTP njenge-substrate, ilandela isimiso sokumatanisa kwesisekelo, futhi ihlanganise umucu owodwa we-DNA ohambisana nesifanekiso se-RNA ohlangothini lwe-5'-3′.I-reverse transcriptase incike ngokuyinhloko kumsebenzi we-DNA polymerase ovela kuzifanekiso ze-DNA noma ze-RNA ngakho-ke awunawo umsebenzi we-3′-5′ we-exonuclease.Kodwa-ke, inomsebenzi we-RNase H, okhawulela ubude be-synthesis be-reverse transcriptase ngokwezinga elithile.Ngenxa yokuthembeka okuphansi kanye nokuqina kwe-wild reverse transcriptase, ososayensi baphinde bayilungisa.

NgobaI-PCRizivivinyo, izinto ezisetshenziswayo eziyinhloko yilezi:ithubhu ye-PCR ngayinye, ishubhu ye-PCR eyi-4/8, amapuleti e-PCR.

I-Labio'sIzinto ezisetshenziswayo ze-PCRube nalokhu okulandelayoizinzuzo:

Amapuleti e-PCR: Ukuhambisana komjikelezo oshisayo obanzi;Ukugqama okuphezulu, ukuhlonza kahle kahle;kahle ukukhanya kwe-fluorescence; kuhleukudluliswa kokushisa;I-DNase eqinisekisiwe, i-RNase, i-DNA, i-PCR inhibitors, kanye ne-pyrogen-free ehloliwe.

Amashubhu e-PCR angawodwana: Ukumelana nokuhwamuka; kuhleukudluliswa kokushisa;ukucaca okuhle kakhulu kokubona; I-DNase eqinisekisiwe, i-RNase, i-DNA, i-PCR inhibitors, kanye ne-pyrogen ehloliwe.

Amashubhu e-PCR angama-4/8: Izindonga ezizacile kakhulu;ukucaca okuphezulu;ukukhanya okuhle kwe-fluorescence;ingasebenzisa embonini yezemithi, imboni yokudla, kanye nebhayoloji yamangqamuzana;ikhwalithi ephezulu, impahla ye-PP emsulwa;I-DNase eqinisekisiwe, i-RNase, i-DNA, i-PCR inhibitors, kanye ne-pyrogen ehloliwe.