Wild Taq DNA Polymerase
Taq DNA Polymerase yog ib qho thermostable DNA polymerase los ntawm Thermus aquaticus YT-1, uas muaj 5′ → 3′ polymerase kev ua haujlwm thiab 5′ flap endonuclease kev ua.
Cheebtsam
| Cheebtsam | HC10 10 A-01 | HC10 10 A-02 | HC10 10 A-03 | HC10 10 A-04 |
| 10 × Taq Buffer | 2 x 1ml | 2 × 10 ml | 2 × 50 ml | 5 × 200 ml |
| DNA Polymerase (5 U / μL) | 0.1ml ib | 1 ml ib | 5 ml ib | 5 × 10 ml |
Kev cia khoom
Kev thauj mus los hauv qab 0 ° C thiab khaws cia ntawm -25 ° C ~ -15 ° C.
Unit txhais
Ib chav tsev yog txhais raws li tus nqi ntawm cov enzyme uas suav nrog 15 nmol ntawm dNTP rau hauv cov khoom siv acid insoluble hauv 30 feeb ntawm 75 ° C.
Kev Tswj Xyuas Zoo
1.Protein Purity Assay (SDS-PAGE):Lub purity ntawm Taq DNA polymerase yog ≥95% txiav txim los ntawm SDS-PAGE tsom xam.
2.Endonuclease kev ua:Yam tsawg kawg ntawm 5 U ntawm Taq DNA polymerase nrog 1 μg λDNA rau 16 teev ntawm 37 ℃ ua rau tsis pom qhov degradation raws li kev txiav txim.
3.Kev Ua Haujlwm Exonuclease:Yam tsawg kawg ntawm 5 U ntawm Taq DNA polymerase nrog 1 μg λ -Hind Ⅲ zom DNA rau 16 teev ntawm 37 ℃ ua rau tsis pom qhov degradation raws li kev txiav txim.
4.Nkaum kev ua si:Yam tsawg kawg ntawm 5 U ntawm Taq DNA polymerase nrog 1 μg pBR322 DNA rau 16 teev ntawm 37 ° C ua rau tsis muaj qhov degradation raws li kev txiav txim.
5.RNase Kev Ua Haujlwm:Yam tsawg kawg ntawm 5 U ntawm Taq DNA polymerase nrog 1.6 μg MS2 RNA rau 16 teev ntawm 37 ° C ua rau tsis muaj qhov degradation raws li kev txiav txim.
6.E. coliDNA:5 U ntawm Taq DNA polymerase yog tshuaj ntsuam xyuas qhov muaj E. coli genomic DNA siv TaqMan qPCR nrog primers tshwj xeeb rau E. coli 16S rRNA locus.E. coli genomic DNA paug yog ≤1 Daim.
7.PCR Amplification (5.0 kb Lambda DNA)- Ib qho 50 µL cov tshuaj tiv thaiv uas muaj 5 ng Lambda DNA nrog 5 units ntawm Taq DNA Polymerase rau 25 lub voj voog ntawm PCR amplification tshwm sim hauv cov khoom xav tau 5.0 kb.
Kev teeb tsa kev daws teeb meem
| Cheebtsam | Ntim |
| Template DNAa | xaiv tau |
| 10 μM Forward Primer | 1 mL ib |
| 10 μM Rov qab Primer | 1 mL ib |
| dNTP Mix (10mM txhua) | 1 mL ib |
| 10 × Taq Buffer | 5l miv |
| Cov DNA Polymeraseb | 0.25 mL ib |
| Cov dej tsis muaj Nuclease | Txog li 50 μL |
Nco tseg:
1) Qhov zoo tshaj plaws tshuaj tiv thaiv concentration ntawm cov qauv sib txawv yog txawv.Cov lus hauv qab no qhia txog kev siv cov qauv kev pom zoo ntawm 50 µL cov tshuaj tiv thaiv kab mob.
| DNA | Tus nqi |
| Genomic | 1 ng - 1 mg |
| Plasmid los yog Viral | 1 pg-1 ib |
2) Qhov zoo tshaj plaws concentration ntawm Taq DNA Polymerase yuav nyob ntawm 0.25 µL ~ 1 µL hauv cov ntawv tshwj xeeb.
Cov tshuaj tiv thaivProgram
| Kauj ruam | Kub(°C) | Sijhawm | Lub voj voog |
| Thawj denaturationa | 95 ℃ | 5 feeb | - |
| Denaturation | 95 ℃ | 15-30 s., kuv | 30-35 Cycle |
| Annealingb | 60 ℃ | 15s ib | |
| Txuas ntxiv | 72 ℃ | 1 kb/min | |
| Kawg Extension | 72 ℃ | 5 feeb | - |
Nco tseg:
1) Thawj qhov xwm txheej denaturation yog tsim rau feem ntau cov tshuaj tiv thaiv amplification thiab tuaj yeem hloov kho raws li qhov nyuaj ntawm cov qauv qauv.Yog tias tus qauv qauv yog qhov nyuaj, lub sijhawm ua ntej denaturation tuaj yeem txuas ntxiv mus rau 5 - 10mins txhawm rau txhim kho qhov pib denaturation.
2) Lub annealing kub yuav tsum tau kho raws li tus nqi Tm ntawm primer, uas feem ntau yog teem rau 3 ~ 5 ℃ qis dua tus nqi Tm ntawm primer;Rau cov qauv nyuaj, nws yog ib qho tsim nyog yuav tsum tau kho qhov kub thiab txias thiab ncua sij hawm ntxiv kom ua tiav kev ua kom muaj zog.
