Wild Taq DNA Polymerase
Taq DNA Polymerase ke DNA polymerase e thermostable e tsoang ho Thermus aquaticus YT-1, e nang le tšebetso ea 5′→3′ polymerase le tšebetso ea 5′ flap endonuclease.
Likaroloana
| Karolo | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10 × Taq Buffer | 2x1 mL | 2 × 10 mL | 2 × 50 mL | 5 × 200 mL |
| Taq DNA Polymerase (5 U/μL) | 0.1 mL | 1 mL | 5 mL | 5 × 10 mL |
Boemo ba polokelo
Lipalangoang ka tlas'a 0°C 'me li bolokoe ho -25°C~-15°C.
Tlhaloso ea Yuniti
Yuniti e le 'ngoe e hlalosoa e le palo ea enzyme e kenyelletsang 15 nmol ea dNTP linthong tse sa keneng tsa asiti ka metsotso e 30 ho 75°C.
Taolo ea Boleng
1.Protein Purity Assay (SDS-PAGE):Bohloeki ba Taq DNA polymerase e ne e le ≥95% e khethiloeng ke tlhahlobo ea SDS-PAGE.
2.EndOnuclease Mosebetsi:Bonyane ba 5 U ea Taq DNA polymerase e nang le 1 μg λDNA bakeng sa lihora tse 16 ho 37 ℃ e etsa hore ho se be le ho senyeha ho ka bonoang joalokaha ho laetsoe.
3.Ts'ebetso ea Exonuclease:Bonyane ba 5 U ea Taq DNA polymerase e nang le 1 μg λ -Hind Ⅲ digest DNA bakeng sa lihora tse 16 ho 37 ℃ e fella ka hore ho se be le ho senyeha ho ka bonoang joalokaha ho laetsoe.
4.Mosebetsi oa Nickase:Bonyane ba 5 U ea Taq DNA polymerase e nang le 1 μg pBR322 DNA bakeng sa lihora tse 16 ho 37 ° C e etsa hore ho se ke ha e-ba le tšenyo e bonahalang e lekanyelitsoeng.
5.Mosebetsi oa RNase:Bonyane ba 5 U ea Taq DNA polymerase e nang le 1.6 μg MS2 RNA bakeng sa lihora tse 16 ho 37 ° C e etsa hore ho se be le ho senyeha ho bonahalang ho ea kamoo ho laetsoeng.
6.E. coliDNA:5 U of Taq DNA polymerase e hlahlojoa boteng ba E. coli genomic DNA e sebelisa TaqMan qPCR e nang le li-primers tse tobileng bakeng sa sebaka sa E. coli 16S rRNA.Tšilafalo ea E. coli genomic DNA ke ≤1 Kopi.
7.Kholiso ea PCR (5.0 kb Lambda DNA)- Karabelo ea 50 µL e nang le 5 ng Lambda DNA e nang le li-unit tse 5 tsa Taq DNA Polymerase bakeng sa li-cycle tse 25 tsa ntlafatso ea PCR e hlahisa sehlahisoa se lebelletsoeng sa 5.0 kb.
Rection Setup
| Likaroloana | Bolumo |
| DNA ea Setšoantšoa | boikhethelo |
| 10 μM Pele Pele | 1 μL |
| 10 μM Reverse Primer | 1 μL |
| dNTP Mix (10mM e 'ngoe le e 'ngoe) | 1 μL |
| 10 × Taq Buffer | 5 μL |
| Taq DNA Polymeraseb | 0.25 μL |
| Metsi a se nang nyutlelie | Ho fihlela ho 50 μL |
Lintlha:
1) Khokahano e nepahetseng ea karabelo ea litempele tse fapaneng e fapane.Tafole e latelang e bonts'a ts'ebeliso e khothaletsoang ea template ea 50 µL reaction system.
| DNA | Chelete |
| Genomic | 1 ng-1 μg |
| Plasmid kapa kokoana-hloko | 1 leq-1 ng |
2) Khokahano e nepahetseng ea Taq DNA Polymerase e ka tloha ho 0.25 µL~1 µL lits'ebetsong tse ikhethileng.
BoitšoaroLenaneo
| Mohato | Mocheso(°C) | Nako | Lipotoloho |
| Denaturation ea pelea | 95 ℃ | 5 mets | - |
| Denaturation | 95 ℃ | 15-30 s | 30-35 Lipotoloho |
| Annealingb | 60 ℃ | 15 s | |
| Katoloso | 72 ℃ | 1kb/mots | |
| Keketso ea ho Qetela | 72 ℃ | 5 mets | - |
Lintlha:
1) Boemo ba pele ba denaturation bo loketse karabelo e ngata ea amplification mme bo ka fetoloa ho latela ho rarahana ha sebopeho sa template.Haeba sebopeho sa template se rarahane, nako ea pre-denaturation e ka atolosoa ho 5 - 10mins ho ntlafatsa phello ea pele ea denaturation.
2) Thempereichara ea annealing e hloka ho lokisoa ho latela boleng ba Tm ba primer, eo ka kakaretso e behiloeng ho 3 ~ 5 ℃ ka tlase ho boleng ba Tm ba primer;Bakeng sa li-templates tse rarahaneng, hoa hlokahala ho fetola mocheso oa annealing le ho eketsa nako ea ho atolosa ho finyella katleho e kholo.
