Hotstart Taq DNA Polymerase
Hot Start Taq DNA Polymerase (Antibody modification) he DNA polymerase thermostable wela wela mai Thermus aquaticus YT-1, nona ka hana polymerase 5′→3′ a me ka hana 5′ flap endonuclease.ʻO ka hoʻomaka wela Taq DNA polymerase he Taq DNA polymerase i hoʻololi ʻia e nā antibodies Taq thermolabile.Hoʻonui ka hoʻololi ʻana i ka antibody i ka kikoʻī, ka naʻau, a me ka hua o ka PCR.
Nā ʻāpana
| ʻāpana | HC1012A-01 | HC1012A-02 | HC1012A-03 | HC1012A-04 |
| 5×HC Taq Buffer | 4×1 mL | 4×10 mL | 4×50 mL | 5×400 mL |
| Hoʻomaka wela Taq DNA Polymerase (Hoʻololi ʻia ka Antibody) (5 U/μL) | 0.1 mL | 1 mL | 5 mL | 10×5 mL |
Nā noi
10 mM Tris-HCl (pH 7.4 ma 25 ℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween20, 0.5% IGEPALCA-630 a me 50% Glycerol.
Kūlana Waihona
ʻO ka halihali ma lalo o 0°C a mālama ʻia ma -25°C~-15°C.
Wehewehe Unite
Hoʻokahi ʻāpana i wehewehe ʻia ʻo ka nui o ka enzyme e hoʻokomo i 15 nmol o dNTP i loko o ka mea hiki ke hoʻoheheʻe ʻia i loko o 30 mau minuke ma 75°C.
Ka Mana Mana
1.Endonuclease Activity:ʻO ka incubation o 20 U o ka enzyme me 4 μg pUC19 DNA no 4 mau hola ma 37 ℃ ka hopena i ʻike ʻole ʻia ka degradation o ka DNA e like me ka hoʻoholo ʻana e ka electrophoresis gel.
2.5 kb Lambda PCR:25 Cycles o ka PCR amplification o 5 ng Lambda DNA me 1.25 units o Taq DNA Polymerase i mua o 200 µM dNTPs a me 0.2 µM primers i ka hopena o ka huahana 5 kb i manaʻo ʻia.
3.Ka hana Exonuclease:ʻO ka hoʻoulu ʻana o kahi hopena 50 µl i loaʻa ka liʻiliʻi o 12.5 U o Taq DNA Polymerase me 10 nmol 5'-FAM oligonucleotide no 30 mau minuke ma 37 ℃ ʻaʻole i ʻike ʻia ka degradation.
4.Hana Hana:ʻO ka hoʻoulu ʻana o 10 µL hopena i loaʻa ka 20 U o ka enzyme me 1μg o RNA transcripts no 2 mau hola ma 37°C ʻaʻole i ʻike ʻia ka hōʻino ʻana o ka RNA e like me ka hoʻoholo ʻana e ka electrophoresis gel.
5.ʻAʻohe wela:ʻAʻole.
Pūnaehana Manaʻo
| Nā ʻāpana | Volume |
| DNA templatea | koho |
| 10 μM Puhi mua | 0.5 μL |
| 10 μM Pumi Huli | 0.5 μL |
| Hui dNTP (10mM kēlā me kēia) | 0.5 μL |
| 5×HC Taq Buffer | 5 μL |
| Taq DNA Polymeraseb(5U/μL) | 0.125 μL |
| Wai nuclease-free | A hiki i ka 25 μL |
Nā memo:
1) a.
| DNA | Ka nui |
| Genomic | 1 ng-1 μg |
| Plasmid a i ʻole Viral | 1 pg-1 ng |
2) b.ʻO ka manaʻo maikaʻi loa o Taq DNA Polymerase mai ka 5-50 units/mL (0.1-0.5 units/25 µL reaction) i nā noi kūikawā.
ʻO ke kaʻa kaʻa kaʻa kaʻa wela
PCR
| ʻanuʻu | Mahana(°C) | Manawa | Nā pōʻaiapuni |
| Denaturation muaa | 95 ℃ | 1-3min | - |
| Denaturation | 95 ℃ | 15-30 s | 30-35 Puka |
| Hoʻopilib | 45-68 ℃ | 15-60 s | |
| Hoʻonui | 68 ℃ | 1kb/min | |
| Hoʻonui hope | 68 ℃ | 5 min | - |
Nā memo:
1) Ua lawa ka denaturation mua o 1 min ma 95°C no ka nui o nā hoʻonui.No nā mamana paʻakikī, ʻoi aku ka lōʻihi o ka denaturation o 2-3mins ma 95 ° C.Me ka colony PCR, ua ʻōlelo ʻia he 5mins denaturation mua ma 95°C.
2) He 15-60 mau kekona ka annealing.Hoʻokumu ʻia ka mahana o Annealing ma ka Tm o ka lua mua a he 45-68 ℃ maʻamau.
