Hotstart Taq DNA Polymerase
Ko te Hot Start Taq DNA Polymerase (te whakarerekētanga a Antibody) he whakamaarama whakamaarama DNA polymerase mai i Thermus aquaticus YT-1, kei a ia he ngohe polymerase 5′→3′ me te ngohe endonuclease 5′ flap.Ko te Taq DNA polymerase wera-tīmata he Taq DNA polymerase i whakarereketia e nga paturopi Taq thermolabile.Ko te whakarereketanga antibody i piki ake te motuhake, te tairongo, me te tuku PCR.
Waehanga
| Waehanga | HC1012A-01 | HC1012A-02 | HC1012A-03 | HC1012A-04 |
| 5×HC Taq Kaipupuri | 4×1 mL | 4×10 mL | 4×50 mL | 5×400 mL |
| Hot Start Taq DNA Polymerase (Antibody whakakē) (5 U/μL) | 0.1 mL | 1 mL | 5 mL | 10×5 mL |
Nga tono
10 mM Tris-HCl (pH 7.4 i te 25 ℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween20, 0.5% IGEPALCA-630 me 50% Glycerol.
Tikanga Rokiroki
Te kawe i raro i te 0°C ka penapena ki te -25°C~-15°C.
Whakamaramatanga Waeine
Kotahi te waeine kua tautuhia ko te nui o te whākōkī e whakauru ana i te 15 nmol o te dNTP ki roto i te waikawa korekore i roto i te 30 meneti i te 75°C.
Mana Kounga
1.EndMahi onuclease:Ko te whakamautanga o te 20 U o te whākōkī me te 4 μg pUC19 DNA mō te 4 hāora i te 37 ℃ karekau e kitea te kino o te DNA i whakatauhia e te electrophoresis gel.
2.5 kb Lambda PCR:25 Nga huringa PCR whakakaha o te 5 ng Lambda DNA me te 1.25 waeine o Taq DNA Polymerase i te aroaro o te 200 µM dNTP me te 0.2 µM tuatahi ka puta te hua 5 kb e tumanakohia ana.
3.Mahi Exonuclease:He werohanga o te tauhohenga 50 µl kei roto te iti rawa o te 12.5 U o Taq DNA Polymerase me te 10 nmol 5'-FAM oligonucleotide mo te 30 meneti i te 37 ℃ karekau e kitea te paheketanga.
4.Mahi RNase:Ko te werohanga o te 10 µL tauhohenga kei roto te 20 U o te enzyme me te 1μg o nga tuhinga RNA mo te 2 haora i te 37°C karekau e kitea te whakahekenga o te RNA i whakatauhia e te electrophoresis gel.
5.Te werawera:Kao.
Pūnaha Tauhohenga
| Waehanga | Rōrahi |
| DNA tauiraa | kōwhiringa |
| 10 μM Tuhinga Whakamua | 0.5 μL |
| 10 μM Tuhinga Whakamuri | 0.5 μL |
| Whakaranu dNTP (10mM ia) | 0.5 μL |
| 5×HC Taq Kaipupuri | 5 μL |
| Taq DNA Polymeraseb(5U/μL) | 0.125 μL |
| Wai kore-nuclease | Tae atu ki te 25 μL |
Tuhipoka:
1) a.
| DNA | Te nui |
| Genomic | 1 ng-1 μg |
| Plasmid, Viral ranei | 1 pg-1 ng |
2) b.Ko te kukū tino pai o te Taq DNA Polymerase mai i te 5-50 wae/mL (0.1-0.5 wae/25 µL tauhohenga) i roto i nga tono motuhake.
Kawa paihikara wera
PCR
| Hipanga | Te pāmahana(°C) | Te wa | Nga huringa |
| Te whakahekenga tuatahia | 95 ℃ | 1-3 meneti | - |
| Te whakahekenga | 95 ℃ | 15-30 s | 30-35 Nga huringa |
| Te whakaeneb | 45-68 ℃ | 15-60 s | |
| Toronga | 68 ℃ | 1kb/min | |
| Toronga Whakamutunga | 68 ℃ | 5min | - |
Tuhipoka:
1) Ko te whakahekenga tuatahi o te 1 min i te 95°C he rawaka mo te nuinga o nga whakakaha.Mo nga tauira uaua, ka tūtohuhia kia roa ake te 2-3 meneti i te 95°C.Ma te PCR koroni, ka tūtohuhia kia 5 meneti te whakaheke i te 95°C.
2) Ko te nuinga o te 15-60 hēkona te mahi whakaene.Ko te paemahana whakaene i runga i te Tm o te takirua tuatahi, he 45-68 ℃ te nuinga.
