FastTaq DNA Polymerase(5'→3' exo-)

Product Information
This product can be used for multiplex PCR and dye-based qPCR reactions, but cannot be used for probe-based qPCR experiments. After receiving this product, please store it immediately in a -20°C constant temperature refrigerator.

Reaction system preparation (25 μL system)
2.5 μL reaction buffer, 0.5-1 μL upstream primer (concentration 10 μM), 0.5-1 μL downstream primer (concentration 10 μM), 0.5 μL probe (concentration 10 μM), 0.4 μL dNTP mix (25 mM each), 0.5-1.25 μL MgCl₂ (100 mM), 0.2-1 μL FastTaq DNA Polymerase (5'-3'exo-) (5 U/μL), 1-5 μL DNA template, add ddH₂O to make up the volume to 25 μL.

Reaction procedure
Pre-denaturation: 95°C, 5 min, 1 cycle;
Denaturation: 95°C, 10 sec, annealing and extension: 60°C, 30 sec, 45 cycles.

On-machine detection
After the reaction system is configured, mix and centrifuge, test on the machine, and analyze the results.

Notes
1. Primer probe and MgCl₂ final concentration can be adjusted according to actual conditions.
2. It is generally recommended to use 95°C, 5 min pre-denaturation. For complex templates, high GC templates or long fragments, it can be extended by 5-10 min.
3. Extension time: When the PCR product is larger than 1kb, the extension time is about 1 minute/kb. When the PCR product is less than 1kb, the extension time is 30-60 sec.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.