Li-enzyme tse 'maloa tse sebelisoang hangata litekong tsa PCR

Polymerase chain reaction, e khutsufalitsoeng joalo kaPCRka Senyesemane, ke mokhoa oa baeloji ea limolek'hule o sebelisoang ho holisa likhechana tse itseng tsa DNA.E ka nkoa e le katiso e khethehileng ea DNA ka ntle ho ’mele, e ka eketsang palo e nyenyane haholo ea DNA haholo.Ka nako eohlePCRkarabelo, sehlopha se le seng sa lintho se phetha karolo ea bohlokoa - li-enzyme.

1. Taq DNA

Litekong matsatsing a pele aPCR, bo-rasaense ba sebelisitse Escherichia coli DNA polymerase I, Empa ho na le bothata ka enzyme ena: e hloka ho tlatsa enzyme e ncha nako le nako ha potoloho e etsoa, ​​​​e leng se etsang hore mehato ea ts'ebetso e be thata hanyenyane 'me ho thata ho ikamahanya ka ho feletseng.Bothata bona bo ile ba rarolloa ka mor'a hore ka phoso bo-rasaense ba arole Taq DNA polymerase ho Thermus aquaticus ka 1988. Ho tloha ka nako eo, ho hōlisa ka mokhoa o itekanetseng oa DNA ho fetohile ntho ea sebele.Ho sibolloa ha enzyme ena ho boetse hoa etsaPCRthekenoloji ke thekenoloji e bonolo, e sebetsang le ea bokahohle.Hajoale, Taq DNA polymerase ke polymerase e atileng haholo ho lisebelisoa tsa DNA.

2. PfuDNA

Joalokaha ho boletsoe ka holimo, Taq DNase e na le kokoana e kholo, kahoo bo-rasaense ba fetotse Taq DNA polymerase ho isa bohōleng bo itseng e le ho qoba ho hōlisa ka tsela e sa tobang ka lebaka la ho se lumellane, ho bakang liphello tse sa nepahalang tsa liteko.Empa phetoho ea Taq DNA polymerase e ka thibela ts'ebetso ea DNA polymerase mocheso oa kamore.PfuDNA polymerase e ka etsa hantle bakeng sa bofokoli bo kaholimo ba Taq DNA polymerase, e le hore karabelo ea PCR e ka etsoa ka mokhoa o tloaelehileng, mme sekhahla sa katleho ea phallo ea liphatsa tsa lefutso e ka ntlafatsoa ka nepo.

3. Reverse Transcriptase

Reverse transcriptase e ile ea sibolloa ka 1970. Enzyme ena e sebelisa RNA e le template, dNTP e le substrate, e latela molao-motheo oa "base pairing", 'me e kopanya DNA e le 'ngoe e tlatsanang le template ea RNA ka tataiso ea 5'-3'.Reverse transcriptase e ipapisitse le ts'ebetso ea DNA polymerase ho tsoa ho litempele tsa DNA kapa tsa RNA, ka hona ha e na ts'ebetso ea 3'-5' exonuclease.Leha ho le joalo, e na le ts'ebetso ea RNase H, e fokotsang bolelele ba motsoako oa reverse transcriptase ho isa bohōleng bo itseng.Ka lebaka la botšepehi bo fokolang le thermostability ea wild reverse transcriptase, bo-rasaense le bona ba e fetotse.

Bakeng saPCRliteko, lisebelisoa tse ka sehloohong ke: tube ea PCR ka bomong, 4/8-strip PCR tube, lipoleiti tsa PCR.

Labio eaLisebelisoa tsa PCRna le tse latelangmelemo:

Lipoleiti tsa PCR: Tšebelisano e pharaletseng ea libaesekele tsa mocheso;Phapang e phahameng, boitsebahatso bo bonolo ba seliba;hantle fluorescence reflection; molemoho fetisa mocheso;DNase e netefalitsoeng, RNase, DNA, PCR inhibitors, le liteko tse se nang pyrogen.

Li-tubes tsa PCR ka bomong: E thibela ho tsoa mouoane;e ntleho fetisa mocheso;e hlakileng haholo; DNase e netefalitsoeng, RNase, DNA, PCR inhibitors, le li-pyrogen tse lekiloeng.

4/8-e hlobolisa li-tubes tsa PCR: Mabota a masesaane haholo; ho hlaka ho hoholo; ponts'o e ntle ea fluorescence;e ka sebelisoa indastering ea meriana, indastering ea lijo, le baeloji ea limolek'hule; thepa ea boleng bo holimo, ea moroetsana ea PP; DNase e netefalitsoeng, RNase, DNA, PCR inhibitors, le li-pyrogen tse lekiloeng.