Hotstart Taq DNA Polymerase
Kupisa Kutanga Taq DNA Polymerase (Antibody gadziriso) inopisa-yekutanga thermostable DNA polymerase kubva kuThermus aquaticus YT-1, ine 5'→ 3′ polymerase chiitiko uye 5' flap endonuclease chiitiko.Iyo inopisa-yekutanga Taq DNA polymerase ndeye Taq DNA polymerase iyo yakagadziridzwa ne thermolabile Taq antibodies.Antibody shanduko yakawedzera kujeka, kunzwa, uye goho rePCR.
Zvikamu
| Chikamu | HC1012A-01 | HC1012A-02 | HC1012A-03 | HC1012A-04 |
| 5 × HC Taq Buffer | 4 × 1 mL | 4 × 10 mL | 4 × 50 mL | 5 × 400 mL |
| Kupisa Kutanga Taq DNA Polymerase (Antibody yakagadziridzwa) (5 U/μL) | 0.1 mL | 1 mL | 5 mL | 10 × 5 mL |
Applications
10 mM Tris-HCl (pH 7.4 pa25℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween20, 0.5% IGEPALCA-630 uye 50% Glycerol.
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.Endonuclease Chiitiko:Kuiswa kwe20 U ye enzyme ine 4 μg pUC19 DNA kwemaawa mana pa37 ℃ kwakaguma nekusaonekwa kushatiswa kweDNA sekutsanangurwa negel electrophoresis.
2.5 kb Lambda PCR:25 Cycles yePCR kukwidziridzwa kwe5 ng Lambda DNA ine 1.25 mayunitsi eTaq DNA Polymerase pamberi pe200 µM dNTPs uye 0.2 µM maprimers anoguma mune inotarisirwa 5 kb chigadzirwa.
3.Exonuclease Chiitiko:Kuiswa kwe50 µl reaction ine hushoma hwe12.5 U yeTaq DNA Polymerase ine 10 nmol 5'-FAM oligonucleotide kwemaminetsi makumi matatu pa37 ℃ haibudisi kusvibiswa kunoonekwa.
4.RNase Chiitiko:Kuiswa kwe10 µL reaction ine 20 U yeenzyme ine 1μg yeRNA zvinyorwa zvemaawa maviri pa37 ° C zvakakonzera kusaonekwa kwekuora kweRNA sekutsanangurwa negel electrophoresis.
5.Kusaita Kupisa:Aihwa.
Reaction System
| Zvikamu | Volume |
| DNA templatea | optional |
| 10 μM Pamberi Pekutanga | 0.5 μL |
| 10 μM Reverse Primer | 0.5 μL |
| dNTP Musanganiswa (10mM imwe neimwe) | 0.5 μL |
| 5 × HC Taq Buffer | 5 μL |
| Taq DNA Polymeraseb(5U/μL) | 0.125 μL |
| Mvura isina nyukliya | Kusvika ku25 μL |
Notes:
1) a.
| DNA | Mari |
| Genomic | 1 ng-1 μg |
| Plasmid kana Viral | 1 p.-1 p |
2) b.Iyo yakakwana yekusangana kweTaq DNA Polymerase inogona kubva pa5-50 mayunitsi/mL (0.1-0.5 mayunitsi/25 µL maitiro) mune akasarudzika maapplication.
Thermal cycling protocol
PCR
| Danho | Tembiricha(°C) | Nguva | Cycles |
| Initial denaturationa | 95 ℃ | 1-3mins | - |
| Denaturation | 95 ℃ | 15-30 p | 30-35 Mitambo |
| Annealingb | 45-68 ℃ | 15-60 s | |
| Extension | 68 ℃ | 1kb/min | |
| Final Extension | 68 ℃ | 5 mins | - |
Notes:
1) Dhinaturation yekutanga ye1 min pa95 ° C inokwana kune akawanda amplifications.Kune ma templates akaoma, kureba denaturation ye2-3mins pa 95 ° C inokurudzirwa.Ne colony PCR, yekutanga 5mins denaturation pa 95 ° C inokurudzirwa.
2) Nhanho yeannealing inowanzoita 15-60 s.Annealing tembiricha yakavakirwa paTm yeprimer pair uye inowanzoita 45-68 ℃.
