Fast Taq DNA polymerase
Cat. No.: SB-G3442-1000U
Size: 1000U
Description
This product, Fast Taq DNA polymerase, is a taq DNA polymerase mutant obtained through in vitro evolutionary screening based on wild-type Taq DNA polymerase. It has 5'-3' polymerization activity and 5'-3' exonuclease activity, no 3'-5' exonuclease activity, the amplification speed is 5-15 s/kb, and the 3' end of the amplification product With "A" base, it can effectively amplify DNA fragments within 10 kb.
- Source : derived from Thermus aquaticus , recombinantly expressed by Escherichia coli.
- Definition of enzyme activity: at 74°C, within 30 minutes, using activated salmon sperm DNA as a template primer, the amount of enzyme required to incorporate 10 nmol of deoxynucleotides into acid-insoluble substances is defined as one enzyme activity unit;
- Purity and concentration: SDS-PAGE detection purity ≥ 95%; endogenous nucleic acid residue < 1 pg/μL (qPCR detection); 5 U/μL.
- 10x Reaction Buffer: 100 mM Tris-HCl, 500 mM KCl, 15 mM MgCl2 , pH 8.3;
- Storage Buffer: 10 mM Tris-HCl, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5% Tween 20, 50% Glycerol, pH 7.4
Storage and transportation
Transport in wet ice packs; store at -20°C, valid for 12 months.
Product Contents
| Component Number |
Component |
G3442-1000U |
| G3442-1 |
Fast Taq DNA polymerase |
200 μL |
| G3442-2 |
10× Reaction buffer |
1mL |
| G3442-3 |
dNTP（10 mM each） |
500 μL |
| Product Manual |
1 copy |
Steps
Commonly used PCR reaction system (50 μL):
| Component |
Volume |
| template |
n (adjustable) |
| Fast Taq DNA polymerase |
0.5 μL (adjustable) |
| dNTP Mix (10 mM each) |
1 μL |
| Forward primer/Reverse primer (10 μM) |
2 μL/2 μL |
| 10× Reaction buffer |
5 μL |
| Nuclease Free Water |
To 50 μL |
Note: a: When plasmid or phage DNA is used as template, the recommended addition amount is 50 ng-5 pg per 50 μL system; when genomic DNA is used as template, the recommended addition amount is 250 ng-50 ng per 50 μL system; cDNA is For the template, it is recommended to dilute 2-100 times, and the amount added should not exceed 10% of the total volume; when the bacterial liquid or crude sample is used as the template, the added amount should not exceed 10% of the reaction volume. Excessive templates can easily lead to non-specific amplification, and too few templates can easily lead to low PCR amplification efficiency. b: The final primer concentration range is 0.2-1.0 μM, and the recommended primer concentration is 0.4 μM. Too few primers may lead to low or no amplification, and excessive primers may lead to non-specific amplification; c: other volumes The reaction system can be adjusted according to the 50 μL reaction system;
Recommended PCR reaction procedure:
| Temperature |
Time |
Cycles |
| 95℃ |
3 min |
1 |
| 95℃ |
5-10 s |
25 - 35 |
| 50℃ - 68℃ |
10-30 s |
| 72℃ |
5-15 s/kb |
| 72℃ |
5-10 min |
1 |
| 4-16°C |
Forever |
|
Precautions
1. The optimization of PCR reaction can be adjusted from the aspects of Mg2+ concentration, template amount, enzyme amount, annealing temperature, etc.
2. Please prepare the reaction system on ice, because Fast Taq DNA polymerase has certain reactivity at room temperature.
3. For your safety and health, please wear a lab coat and disposable gloves for operation.
