Wild Taq DNA Polymerase
Taq DNA Polymerase ndeye thermostable DNA polymerase kubva kuThermus aquaticus YT-1, ine 5'→3' polymerase chiitiko uye 5' flap endonuclease chiitiko.
Zvikamu
| Chikamu | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10 × Taq Buffer | 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 |
Storage Condition
Kutakura pasi pe 0°C uye kuchengetwa pa -25°C~-15°C.
Chikamu Tsanangudzo
Imwe yuniti inotsanangurwa sehuwandu hwe enzyme inobatanidza 15 nmol ye dNTP mu acid isinganyunguriki zvinhu mumaminitsi makumi matatu pa 75 ° C.
Quality Control
1.Protein Purity Assay (SDS-PEJI):Kuchena kweTaq DNA polymerase yaive ≥95% yakatarwa neSDS-PAGE ongororo.
2.Endonuclease Chiitiko:Hushoma hwe5 U yeTaq DNA polymerase ine 1 μg λDNA kwemaawa gumi nematanhatu pa37 ℃ inoguma nekusaona kushatisa sekunge kwakatemwa.
3.Exonuclease Chiitiko:Iri shoma ye5 U yeTaq DNA polymerase ine 1 μg λ -Hind Ⅲ digest DNA kwemaawa gumi nematanhatu pa37 ℃ inokonzeresa kuparara kunoonekwa sekurongwa.
4.Nickase Chiitiko:Hushoma hwe5 U yeTaq DNA polymerase ine 1 μg pBR322 DNA kwemaawa gumi nematanhatu pa37°C inokonzeresa kusadzikira kunooneka sekunge kwatemwa.
5.RNase Chiitiko:Hushoma hwe5 U yeTaq DNA polymerase ine 1.6 μg MS2 RNA kwemaawa gumi nematanhatu pa37°C inoguma pasina kushatisa kunoonekwa sekurongwa.
6.E. coliDNA:5 U yeTaq DNA polymerase inoongororwa kuvapo kweE. coli genomic DNA uchishandisa TaqMan qPCR ine maprimers chaiwo eE. coli 16S rRNA locus.Iyo E. coli genomic DNA kusvibiswa ndeye ≤1 Copy.
7.PCR Amplification (5.0 kb Lambda DNA)-A 50 µL reaction ine 5 ng Lambda DNA ine 5 mayunitsi eTaq DNA Polymerase yemakumi maviri neshanu kutenderera kwePCR amplification inoguma mune inotarisirwa 5.0 kb chigadzirwa.
Reaction Setup
| Zvikamu | Volume |
| DNA templatea | optional |
| 10 μM Pamberi Pekutanga | 1 μL |
| 10 μM Reverse Primer | 1 μL |
| dNTP Musanganiswa (10mM imwe neimwe) | 1 μL |
| 10 × Taq Buffer | 5 μL |
| Taq DNA Polymeraseb | 0.25 μL |
| Mvura isina nyukliya | Kusvika ku50 μL |
Notes:
1) Iyo yakakwana maitiro ekusangana kweakasiyana matemplate akasiyana.Tafura inotevera inoratidza yakakurudzirwa template kushandiswa kwe50 µL reaction system.
| DNA | Mari |
| Genomic | 1 ng-1 μg |
| Plasmid kana Viral | 1 p.-1 p |
2) Iyo yakakwana yekusangana kweTaq DNA Polymerase inogona kubva ku0.25 µL ~ 1 µL mumashandisirwo akasarudzika.
ReactionChirongwa
| Danho | Tembiricha(°C) | Nguva | Cycles |
| Initial denaturationa | 95 ℃ | 5 mins | - |
| Denaturation | 95 ℃ | 15-30 p | 30-35 Mitambo |
| Annealingb | 60 ℃ | 15 p | |
| Extension | 72 ℃ | 1kb/min | |
| Final Extension | 72 ℃ | 5 mins | - |
Notes:
1) Iyo yekutanga denaturation mamiriro akakodzera kune akawanda amplification maitiro uye anogona kugadziridzwa zvichienderana nekuoma kwe template chimiro.Kana iyo template dhizaini yakaoma, iyo pre-denaturation nguva inogona kuwedzerwa kusvika ku5 - 10mins kuvandudza iyo yekutanga denaturation mhedzisiro.
2) Iyo tembiricha yeannealing inoda kugadziridzwa zvinoenderana neTm kukosha kweiyo primer, iyo inowanzoiswa ku3 ~ 5 ℃ yakaderera pane Tm kukosha kweiyo primer;Kune matemplate akaoma, zvinodikanwa kugadzirisa tembiricha yeannealing uye kuwedzera nguva yekuwedzera kuti uwane kukwidziridzwa kwakanaka.
