Wild Taq DNA Polymerase
I-Taq DNA Polymerase yi-polymerase ye-DNA ye-thermostable esuka kwi-Thermus aquaticus YT-1, enomsebenzi we-5′→3′ polymerase kunye ne-5′ flap endonuclease.
Amacandelo
| Icandelo | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10× Taq Buffer | 2×1 mL | 2×10 mL | 2×50 mL | 5×200 mL |
| I-Taq DNA Polymerase (5 U/μL) | 0.1 ml | 1 ml | 5 ml | 5×10 mL |
Imeko yoGcino
Ukuthutha ngaphantsi kwe-0°C kwaye igcinwe kwi-25°C~-15°C.
Inkcazelo yeyunithi
Iyunithi enye ichazwa njengobungakanani be-enzyme edibanisa i-15 nmol ye-dNTP kwizinto ezingenakunyibilika ze-asidi kwimizuzu engama-30 kwi-75 ° C.
Ulawulo lwemeko
1.Uvavanyo lweProtein Purity Assay (SDS-PAGE):Ukucoceka kwe-polymerase ye-Taq DNA yayingu-≥95% enqunywe nguhlalutyo lwe-SDS-PAGE.
2.EndOnuclease Umsebenzi:Ubuncinci be-5 U ye-Taq DNA polymerase ene-1 μg λDNA kwiiyure ze-16 kwi-37 ℃ ibangela ukuba kungabikho ukuthotywa okubonakalayo njengoko kumisiwe.
3.Umsebenzi we-Exonuclease:Ubuncinci be-5 U ye-Taq DNA polymerase ene-1 μg λ -Hind Ⅲ digest DNA iiyure ezili-16 kuma-37 ℃ kubangela ukuba kungabikho ukuthotywa okubonakalayo njengoko kumisiwe.
4.Umsebenzi we-Nickase:Ubuncinci be-5 U ye-Taq DNA polymerase ene-1 μg pBR322 DNA kwiiyure ze-16 kwi-37 ° C ibangela ukuba kungabikho ukuthotywa okubonakalayo njengoko kumisiwe.
5.Umsebenzi we-RNase:Ubuncinci be-5 U ye-Taq DNA polymerase ene-1.6 μg MS2 RNA kwiiyure ze-16 kwi-37 ° C ibangela ukuba kungabikho ukuthotywa okubonakalayo njengoko kumisiwe.
6.E. coliI-DNA:I-5 U ye-Taq DNA polymerase ihlolwe ubukho be-E. coli genomic DNA usebenzisa i-TaqMan qPCR kunye ne-primers ekhethekileyo ye-E. coli 16S rRNA locus.Ungcoliseko lwe-E. coli genomic DNA yi ≤1 Ikopi.
7.Ukwandiswa kwePCR (5.0 kb Lambda DNA)-Impendulo ye-50 µL equlethe i-5 ng Lambda DNA eneyunithi ezi-5 ze-Taq DNA Polymerase kwimijikelo engama-25 yeziphumo zokukhulisa i-PCR kwimveliso elindelekileyo ye-5.0 kb.
Ukuseta iRection
| Amacandelo | Umthamo |
| Isakhelo seDNAa | ngokuzikhethela |
| 10 μM Phambili iPrimer | 1 μL |
| 10 μM Ukubuyisela umva iPrimer | 1 μL |
| I-dNTP Mix (10mM nganye) | 1 μL |
| 10×Taq Buffer | 5 μL |
| Taq DNA Polymeraseb | 0.25 μL |
| Amanzi angenanyukliya | Ukuya kuthi ga kwi-50 μL |
Amanqaku:
I-1) Ugxininiso oluchanekileyo lokusabela kwiitemplates ezahlukeneyo zahlukile.Le theyibhile ilandelayo ibonisa usetyenziso olucetyiswayo lwetemplate ye-50 µL yesixokelelwano sokusabela.
| DNA | Isixa |
| Genomic | 1 ng-1 μg |
| I-Plasmid okanye iViral | 1 iphe-1 ng |
2) Olona xinzelelo luphezulu lwe-Taq DNA Polymerase lunokusuka kwi-0.25 µL~1 µL kwizicelo ezikhethekileyo.
UkusabelaInkqubo
| Inyathelo | Ubushushu(°C) | Ixesha | Imijikelo |
| I-denaturation yokuqalaa | 95 ℃ | 5 imiz | - |
| I-Denaturation | 95 ℃ | 15-30 s | 30-35 Imijikelezo |
| Ukuhlaziyab | 60 ℃ | 15 s | |
| Ulwandiso | 72 ℃ | 1kb/min | |
| Ukwandiswa kokugqibela | 72 ℃ | 5 imiz | - |
Amanqaku:
I-1) Imeko yokuqala ye-denaturation ifanelekile kwiimpendulo ezininzi ze-amplification kwaye inokuguqulwa ngokobunzima besakhiwo setemplate.Ukuba ulwakhiwo lwethemplate luyinkimbinkimbi, ixesha lokuchaza kwangaphambili linganwetshwa ukuya kwi-5 - 10mins ukuphucula umphumo wokuqala we-denaturation.
2) Ukushisa kwe-annealing kufuna ukulungiswa ngokwexabiso le-Tm le-primer, elibekwe ngokubanzi kwi-3 ~ 5 ℃ ngaphantsi kwexabiso le-Tm le-primer;Kwiitemplates eziyinkimbinkimbi, kuyimfuneko ukulungelelanisa ubushushu be-annealing kunye nokwandisa ixesha lokwandisa ukufezekisa ukukhulisa kakuhle.
