| Source : |
E.coli |
| Tag : |
Non |
| Description : |
Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activi and a 5´ flap endonuclease activity. Taq 5X Master Mix is an optimized ready-to-use solution containing Taq DNA Polymerase, dNTPs, MgCl2, KCl and stabilizers. It is best for routine PCR applications from templates including pure DNA solutions, bacterial colonies, and cDNA. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA. |
| ReagentsSupplied : |
MgCl2 (25 mM) |
| 1X Taq Master Mix : |
10 mM Tris-HCl (pH 8.6 @ 25°C); 50 mM KCl; 1.5 mM MgCl2; 25 units/ml Taq DNA Polymerase; 0.2 mM dNTPs each; 5% glycerol; 0.08% Ethoxylated tert-octylphenol; 0.05% Tween-20 |
| Applications : |
PCR; Primer Extension; Microarray Analysis; High-Throughput PCR |
| Heat Inactivation : |
No |
| Unit Definition : |
One unit of Taq DNA Polymerase is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C. |
| Unit AssayConditions : |
1X ThermoPol Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA. |
| ReactionConditions : |
1X Taq Master Mix, DNA template and primers in a total reaction volume of 25 or 50 µl. |
| GeneralNotes : |
Taq 5X Master Mix is stable for fifteen freeze-thaw cycles when stored at -20°C. Taq 5X Master Mix is also stable for six months at 4°C, so for frequent use, an aliquot may be kept at 4°C. We recommend assembling all reaction components on ice and quickly transferring the reactions to a thermocycler preheated to the denaturation temperature (95°C). |
| Reaction Setup : |
| Component | 25 µl Reaction | 50 µl Reaction | Final Concentration | | 10 µM Forward Primer | 0.5 µl | 1 µl | 0.2 µM (0.05–1 µM) | | 10 µM Reverse Primer | 0.5 µl | 1 µl | 0.2 µM (0.05–1 µM) | | Template DNA | variable | variable | <1,000 ng | | Taq 5X Master Mix | 5 µl | 10 µl | 1X | | Nuclease-free water | to 25 µl | to 50 µl | | | Step | | | Denaturation | | | 30cycles | | | 45–68°C | 15-60 seconds | | 68°C | 1 minute per kb | | Final extension | | | Hold | 4–10°C | |
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