Wild Taq DNA Polymerase
Taq DNA Polymerase bụ DNA polymerase nke na-ekpo ọkụ sitere na Thermus aquaticus YT-1, nke nwere ọrụ 5′→3′ polymerase na ọrụ 5′ flap endonuclease.
Ngwa
| Akụkụ | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| Ihe nchekwa 10× Taq | 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 |
Ọnọdụ Nchekwa
Ụgbọ njem n'okpuru 0 Celsius ma debe ya na -25C ~ -15C.
Nkọwa nkeji
A kọwapụtara otu nkeji dị ka ọnụọgụ enzyme nke na-etinye 15 nmol nke dNTP n'ime ihe na-adịghị edozi acid n'ime nkeji 30 na 75°C.
Njikwa ogo
1.Nnwale ịdị ọcha nke Protein (SDS-PAGE):Ịdị ọcha nke Taq DNA polymerase bụ ≥95% kpebisiri ike site na nyocha SDS-PAGE.
2.Endonuclease Ọrụ:Opekempe 5 U nke Taq DNA polymerase nwere 1 μg λDNA maka awa 16 na 37 ℃ na-arụpụta enweghị mmebi a na-achọpụta dị ka ekpebisiri ike.
3.Ọrụ Exonuclease:Opekempe nke 5 U nke Taq DNA polymerase nwere 1 μg λ -Hind Ⅲ mgbari DNA maka awa 16 na 37 ℃ na-arụpụta enweghị mmebi a na-achọpụta dị ka ekpebisiri ike.
4.Ọrụ Nickase:Opekempe nke 5 U nke Taq DNA polymerase nwere 1 μg pBR322 DNA maka awa 16 na 37ºC na-arụpụta enweghị mmebi a na-achọpụta dị ka ekpebisiri ike.
5.Ọrụ RNase:Opekempe 5 U nke Taq DNA polymerase nwere 1.6 μg MS2 RNA maka awa 16 na 37°C na-arụpụta enweghị mmebi nke enwere ike ịchọpụta dị ka ekpebisiri ike.
6.E. coliDNA:A na-enyocha 5 U of Taq DNA polymerase maka ọnụnọ nke E. coli genomic DNA na-eji TaqMan qPCR na primers kpọmkwem maka ebe E. coli 16S rRNA.Ihe mmetọ DNA nke E.coli genomic bụ ≤1 Detuo.
7.PCR Amplification (5.0kb Lambda DNA)- Mmeghachi omume 50µL nwere 5 ng Lambda DNA nwere nkeji 5 nke Taq DNA Polymerase maka usoro 25 nke nsonaazụ PCR na-atụ anya na ngwaahịa 5.0 kb.
Ntọala mmeghachi omume
| Ngwa | Olu |
| DNA templatea | nhọrọ |
| 10 μM na-aga n'ihu | 1 μl |
| 10 μM Reverse Primer | 1 μl |
| Ngwakọta dNTP (10mM nke ọ bụla) | 1 μl |
| Ihe nchekwa 10 × Taq | 5 μl |
| DNA Polymeraseb | 0.25 μl |
| Mmiri na-enweghị nuklia | Ihe dị ka 50 μl |
ndetu:
1) Mmetụta mmeghachi omume kachasị mma nke ndebiri dị iche iche dị iche iche.Tebụlụ na-esote na-egosi ojiji nke usoro mmeghachi omume 50µL akwadoro.
| DNA | Ọnụ ego |
| Genomic | 1ng-1 μg |
| Plasmid ma ọ bụ Viral | 1 pg-1 nkp |
2) Ntụnye kachasị mma nke Taq DNA Polymerase nwere ike ịdị na 0.25 µL ~ 1 µL na ngwa pụrụ iche.
Mmeghachi omumeMmemme
| Nzọụkwụ | Okpomọkụ(Celsius C) | Oge | okirikiri |
| denaturation nke mbụa | 95 ℃ | 5mins | - |
| Denaturation | 95 ℃ | 15-30 nkeji | Usoro 30-35 |
| Na-ewe iweb | 60 ℃ | 15 s | |
| Mgbatị | 72 ℃ | 1kb/min | |
| Mgbatị ikpeazụ | 72 ℃ | 5mins | - |
ndetu:
1) The mbụ denaturation ọnọdụ kwesịrị ekwesị maka ọtụtụ mmụba mmeghachi omume na ike gbanwee dị ka mgbagwoju nke template Ọdịdị.Ọ bụrụ na usoro ndebiri dị mgbagwoju anya, enwere ike ịgbatị oge tupu denaturation ruo 5 - 10mins iji melite mmetụta denaturation mbụ.
2) The annealing okpomọkụ kwesịrị ịgbanwe dị ka Tm uru nke primer, nke a na-emekarị ka 3 ~ 5 ℃ dị ala karịa Tm uru nke primer;Maka ndebiri mgbagwoju anya, ọ dị mkpa iji dozie okpomọkụ na-ekpo ọkụ na ịgbatị oge mgbatị iji nweta nkwalite dị mma.
