Wild Taq DNA Polymerase
Ko te Taq DNA Polymerase he DNA polymerase whakamaarama mai i Thermus aquaticus YT-1, kei a ia he ngohe polymerase 5′→3′ me te ngohe 5′ flap endonuclease.
Waehanga
| Waehanga | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10× Taq Kaipupuri | 2×1 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 |
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.Te Whakatairanga Maamaa Protein (SDS-PAGE):Ko te ma o Taq DNA polymerase i ≥95% i whakatauhia e te tātaritanga SDS-PAGE.
2.EndMahi onuclease:Ko te iti rawa o te 5 U o te Taq DNA polymerase me te 1 μg λDNA mo nga haora 16 i te 37 ℃ ka kore e kitea te paheketanga kua whakatauhia.
3.Mahi Exonuclease:Ko te iti rawa o te 5 U o te Taq DNA polymerase me te 1 μg λ -Hind Ⅲ digest DNA mo nga haora 16 i te 37 ℃ ka kore e kitea te paheketanga ka kitea.
4.Mahi Nickase:Ko te iti rawa o te 5 U o te Taq DNA polymerase me te 1 μg pBR322 DNA mo te 16 haora i te 37°C ka kore e kitea te paheketanga kua whakatauhia.
5.Mahi RNase:Ko te iti rawa o te 5 U o te Taq DNA polymerase me te 1.6 μg MS2 RNA mo te 16 haora i te 37°C ka kore e kitea te paheketanga kua whakatauhia.
6.E. coliDNA:Ko te 5 U o Taq DNA polymerase ka tirotirohia mo te aroaro o te E. coli genomic DNA ma te whakamahi i te TaqMan qPCR me nga tohu tuatahi mo te waahi E. coli 16S rRNA.Ko te pokenga DNA genomic E. coli he ≤1 Tārua.
7.Whakanui PCR (5.0 kb Lambda DNA)- He tauhohenga 50 µL kei roto te 5 ng Lambda DNA me te 5 waeine o Taq DNA Polymerase mo nga huringa 25 o te whakakaha PCR ka puta te hua 5.0 kb e tumanakohia ana.
Tauhohenga Tatūnga
| Waehanga | Rōrahi |
| DNA tauiraa | kōwhiringa |
| 10 μM Tuhinga Whakamua | 1 μL |
| 10 μM Tuhinga Whakamuri | 1 μL |
| Whakaranu dNTP (10mM ia) | 1 μL |
| 10×Taq Kaipupuri | 5 μL |
| Taq DNA Polymeraseb | 0.25 μL |
| Wai kore-nuclease | Tae atu ki te 50 μL |
Tuhipoka:
1) He rereke te kukū tauhohenga pai o nga tauira rereke.Ko te ripanga e whai ake nei e whakaatu ana i te whakamahi tauira tauira 50 µL pūnaha tauhohenga.
| DNA | Te nui |
| Genomic | 1 ng-1 μg |
| Plasmid, Viral ranei | 1 pg-1 ng |
2) Ko te kukū tino pai o te Taq DNA Polymerase mai i te 0.25 µL~1 µL i roto i nga tono motuhake.
TauhohengaPapatono
| Hipanga | Te pāmahana(°C) | Te wa | Nga huringa |
| Te whakahekenga tuatahia | 95 ℃ | 5min | - |
| Te whakahekenga | 95 ℃ | 15-30 s | 30-35 Nga huringa |
| Te whakaeneb | 60 ℃ | 15 s | |
| Toronga | 72 ℃ | 1kb/min | |
| Toronga Whakamutunga | 72 ℃ | 5min | - |
Tuhipoka:
1) He pai te ahua o te denaturation tuatahi mo te nuinga o nga tauhohenga whakakaha, ka taea te whakarereke i runga i te uaua o te hanganga tauira.Mena he uaua te hanganga tauira, ka taea te whakaroa ki te 5 - 10 meneti hei whakapai ake i te paanga o te denaturation tuatahi.
2) Me whakatika te pāmahana o te annealing kia rite ki te uara Tm o te tuatahi, e whakaritea ana ki te 3~5 ℃ iti iho i te uara Tm o te tuatahi;Mo nga tauira uaua, he mea tika ki te whakatika i te pāmahana whakahiato me te whakaroa i te wa toronga kia pai ai te whakakaha.
