Taq DNA Polymerase
Cat. No.: SB-G3441-1000U
Size: 1000U
Description
The Taq DNA polymerase catalyzes 5´→3´synthesis of DNA, which can efficiently amplify DNA fragments up to 7 kb. The enzyme also exhibits 3'→5' exonuclease (proofreading) activity without 5'→3' exonuclease activity. The 3' end of the PCR product has an "A" base.
- Applications: Ideal for routine PCR applications, TA cloning
- Source : E.coli cells with a cloned polA gene from Thermus aquaticus.
- Concentration: 5 U/μL
- Endogenous nucleic acid: ＜1 pg/μL by qPCR
- Definition of Activity Unit : One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into an acid-insoluble substance 30 min at 74 °C
- Purity: ≥95% by SDS-PAGE
- 10x Reaction Buffer: 100 mM Tris-HCl，500 mM KCl，15 mM MgCl2，pH 8.3
- Storage Buffer: 10 mM Tris-HCl (pH 7.4)，100 mM KCl，1 mM DTT，0.1 mM EDTA，0.5% Tween 20，50% (v/v) Glycerol
- Storage Conditions: Store at –20°C up to 12 months.
Product Contents
| Component Number |
Component |
G3441-1000U |
| G3441-1 |
Taq DNA polymerase |
200 μL |
| G3441-2 |
10× Reaction buffer |
1mL |
| G3441-3 |
dNTP（10 mM each） |
500 μL |
| Product Manual |
1 copy |
Assay Protocol
Routine PCR Protocol
1. Reaction setup:
We recommend assembling all reaction components on ice and quickly transferring the reactions to a thermocycler preheated to the denaturation temperature (95°C).
| Component |
Volume |
| 10× Reaction buffer |
5 μL |
| Taq DNA polymerase |
0.5 μL (variable) |
| dNTP Mix (10 mM each) |
1 μL |
| Forward primer/Reverse primer (10 μM) |
2 μL/2 μL |
| Template |
Variable (0.1-50 ng) |
| Nuclease Free Water |
To 50 μL |
It could be added to no more than 10% DMSO to the reaction system for high GC templates.
The reaction volumes can be scaled up or down providing that the final concentrations of the components (DNA, dNTPs, labeled dNTP) are as indicated in the protocol.
2. Gently vortex the samples and spin down.
3. Perform PCR using the recommended thermal cycling conditions outlined below:
| Step |
Temperature |
Time |
Number of Cycles |
| Initial Denaturation |
95℃ |
3 min |
1 |
| Denaturation |
95℃ |
15-30 s |
25 - 35 |
| Annealing |
50℃ - 68℃ |
15-30 s |
| Extension |
72℃ |
1 min/kb |
| Final extension |
72℃ |
5-10 min |
1 |
Note
1. It is recommended that the reaction system should be prepared on ice.
2. The optimization of PCR reactions can be adjusted from Mg
2+ concentration, template amount, enzyme amount, annealing temperature, etc.
3. For your safety and health, please wear safety glasses, gloves, or protective clothing.
