Hotstart Taq DNA Polymerase
Hot Committitur Taq DNA Polymerase (anticorporis modificatio) est thermostici DNA polymerasis calidus e Thermo aquatico YT-1，, qui polymerasium habet 5′ →3′ activitatem 5´ moto endonuclease.Polymerasium calidum Taq DNA est Polymerasis Taq DNA quae ab elementorum thermolabili Taq modificatur.Anticorporis modificatio aucta specificitate, sensibilitate, ac cede PER.
Components
| Component | HC1012A-01 | HC1012A-02 | HC1012A-03 | HC1012A-04 |
| 5×HC Taq Buffer | 4×1 mL | 4×10 mL | 4×50 mL | 5×400 mL |
| Hot Committitur Taq DNA Polymerase (Anticorpus mutatio) (5 U/μL) | 0.1 mL | 1 mL | 5 mL | 10×5 mL |
Applications
10 mM Tris-HCl (pH 7.4 ad 25℃), 100 mM KCl, 0,1 mM EDTA, 1 mM dithiothreitolum, 0.5% Tween20, 0.5% IGEPALCA-630 et 50% Glycerolum.
Repono Condition
Transportatio sub 0°C et reponenda -25°C~-15°C.
Unitas Definition
Una unitas definitur quantum enzyme, quod 15 nmol de dNTP in acidi insolubili materia in 30 minuta ad 75°C incorporate definitur.
Regimen quālitātis
1.Endonuclease Actio:Incubatio 20 U enzyme cum 4 μg pUC19 DNA pro 4 horis 37℃ consecuta est nullo detectabili degradatione DNA secundum electronicas electrophoresis determinatae.
2.5 kb Labda pcr:25 Cycles PCR amplificationis 5 ng Lambda DNA cum 1.25 unitatibus Taq DNA Polymerase coram 200 µM dNTPs et 0.2 µM primorum eventus in producto exspectato 5 kb.
3.Exonuclease Actio:Incubatio 50 μl reactionis continens minimum 12.5 U Taq DNA Polymerase cum 10 nmol 5´-FAM oligonucleotide per 30 minuta ad 37℃ degradationem detectam nullam praebet.
4.RNase Actio:Incubatio 10 µL reactionem continens XX U enzyme cum 1μg RNA transcriptorum per 2 horas ad 37°C consecuta est nullo detectabili deformitate RNA iuxta electronicam electrophoresis determinatam.
5.Calor Inactivation:Nec.
Ratio reactionem
| Components | Magnitudo |
| Formula DNAa | ad libitum |
| 10 μM Primus | 0.5 μL |
| 10 μM Reverse Primer | 0.5 μL |
| dNTP Mix (10mM each) | 0.5 μL |
| 5×HC Taq Buffer | 5 μL |
| Taq DNA Polymeraseb(5U/μL) | 0.125 μL |
| Nucleas-liber aqua | Usque ad 25 μL |
Notae:
1) a.
| DNA | Moles |
| Genomic | 1 ng-1 μg |
| Plasmid vel Virales | 1 pg-1 ng |
2) b.Optima intentio Taq DNA Polymerase ab unitates 5-50 vagari potest / mL (0.1-0.5 unitates/25 µL reactiones) in applicationibus specialibus.
Scelerisque revolutio protocol
PCR
| Gradus | Temperature(°C) | Tempus | Cycles |
| Coepi denaturationa | 95 | 1-3mins | - |
| Denaturation | 95 | 15-30 s | 30-35 Cycli |
| Annealingb | 45-68 | 15-60 s | |
| Extensio | 68 | 1kb/min | |
| Extensio finalis | 68 | 5mins | - |
Notae:
1) Denaturatio initialis 1 min ad 95°C satis est ad plurimas amplificationes.Pro difficilibus exemplaribus, longior denaturatio 2-3minum in 95°C commendatur.Colonia PCR, 5 mins denaturation initialis in 95°C commendatur.
2) In furnum gradus est proprie 15-60 s.Temperatura annalium in Tim primi paris fundatur et est typice 45-68℃.
