Ma enzymes akati wandei anowanzo shandiswa muPCR kuyedza

Polymerase chain reaction, yakapfupikiswa sePCRmuChirungu, inyanzvi yebhayoroji yemolecular inoshandiswa kukudza zvimedu zveDNA.Inogona kutaridzirwa seyakakosha DNA replication kunze kwemuviri, iyo inogona kuwedzera zvakanyanya huwandu hushoma hweDNA.Munguva yosePCRmaitiro ekuita, imwe kirasi yezvinhu inoita basa rakakosha - enzymes.

1. Taq DNA

Mukuedza mumazuva ekutanga ePCR, masayendisiti akashandisa Escherichia coli DNA polymerase I, Asi pane dambudziko ne enzyme iyi: inoda kuzadza enzyme itsva nguva dzose kana kutenderera kunoitwa, izvo zvinoita kuti matanho ekuvhiya ave akaoma uye zvakaoma kunyatsozvikudza.Dambudziko iri rakagadziriswa mushure mekunge masayendisiti atsaura netsaona Taq DNA polymerase kubva kuThermus aquaticus muna 1988. Kubva ipapo, iyo otomatiki amplification yeDNA yave kuitika.Kuwanikwa kwe enzyme iyi kunoitawoPCRtekinoroji iri nyore, inoshanda uye yepasirese tekinoroji.Parizvino, Taq DNA polymerase ndiyo yakajairika polymerase muDNA kits.

2. PfuDNA

Sezvambotaurwa pamusoro apa, Taq DNase ine tsikidzi huru, saka masayendisiti akashandura Taq DNA polymerase kune imwe nhanho kuitira kuti adzivise kukwidziridzwa kusiri kwakanangana nekuda kwekusawirirana, zvichikonzera mhedzisiro yebvunzo isiriyo.Asi kugadziridzwa kweTaq DNA polymerase kunogona kutadzisa kuita kweDNA polymerase pakudziya kwekamuri.PfuDNA polymerase inogona kunyatso gadzirisa izvo zviri pamusoro zvakaipa zveTaq DNA polymerase, kuitira kuti PCR reaction iitwe zvakajairwa, uye mwero wekubudirira wechinangwa chekusimudzira gene unogona kuvandudzwa.

3. Reverse Transcriptase

Reverse transcriptase yakawanikwa muna 1970. Iyi enzyme inoshandisa RNA se template, dNTP se substrate, inotevera musimboti we base pairing, uye inogadzirisa DNA imwe strand inopindirana neRNA template mu 5'-3' nzira.Reverse transcriptase inonyanya kutsamira pane DNA polymerase chiitiko kubva kuDNA kana RNA matemplate uye saka haina 3'-5′ exonuclease chiitiko.Nekudaro, ine RNase H chiitiko, iyo inodzika iyo synthesis kureba kwereverse transcriptase kune imwe nhanho.Nekuda kwekuderera kwekutendeseka uye kudziya kwemusango reverse transcriptase, vesainzi vakaigadzirisa zvakare.

ForPCRkuyedza, izvo zvinodyiwa zvakanyanya ndeizvi: yemunhu PCR chubhu, 4/8-strip PCR chubhu, PCR mahwendefa.

Labio'sPCR inoshandiswaiva nezvinoteverazvakanakira:

PCR mahwendefa: Broad thermal cycler kuenderana;High-kusiyana, nyore tsime kuzivikanwa;zvakanaka fluorescence reflection; zvakanakakupisa kupisa;Certified DNase, RNase, DNA, PCR inhibitors, uye yakaedzwa isina pyrogen.

Mumwe nomumwe PCR tubes: Evaporation-resistant;zvakanakakupisa kupisa;yakanakisa kujeka kwemaziso; Certified DNase, RNase, DNA, PCR inhibitors, uye yakaedzwa isina pyrogen.

4/8-inobvisa PCR machubhu: Ultra-mutete madziro; kujeka kwepamusoro; yakanaka fluorescence kuratidzwa;inogona kushandisa muindasitiri yemishonga, indasitiri yechikafu, uye molecular biology; yepamusoro-soro, mhandara PP zvinhu; Certified DNase, RNase, DNA, PCR inhibitors, uye yakaedzwa pyrogen-isina.