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2x Taq PCR Premix

 

Description: 2x Taq PCR Premix is an ready-to-use optimized solution containing Taq DNA polymerase, standard Taq reaction buffer, dNTPs, tracking dyes and stabilizers. It is ideally suited to routine PCR applications such as sub-cloning, colony screening and genotyping. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA. Blue dye migrates approximately at 4 kb, and yellow dye migrates approximately at 50 bp.

Source: Recombinant Taq DNA polymerase purified from an E. coli strain carrying Thermus aquaticus gene.Taq DNA polymerase is a thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a double-strand specific 5´→3´ exonuclease activity. TP01-01 contains 250 units of Taq DNA polymerase. TP01-02 contains 1000 units of Taq DNA polymerase.

Applications

  • PCR
  • Primer Extension
  • Microarray Analysis
  • High-Throughput PCR

Storage Condition: 2x TaqPCR Premix should be stored at -20oC. Limited (up to 10 times) freeze-thaw does not affect PCR performance. For daily use, it’s recommended to keep an aliquot at 4°C, which is stable up to 6 weeks.

Protocol
These recommendations serve as a starting point. In order to maximize amplification, the reaction conditions may require optimization.
  1. Thaw 2xTaq PCR Premix at room temperature and mix well by inverting several times before use.
  2. Prepare the following reaction in a thin-walled PCR tube on ice:
Component
25 µl
50 µl
Final
2x Taq PCR Premix
12.5 µl
25 µl
1x
5’ Primer (10 µM)
0.5 µl
1 µl
0.2 µM
3’ Primer (10 µM)
0.5 µl
1 µl
0.2 µM
DNA template
Determined by user
0.1–1ng/ml plasmid DNA
1-10ng/ml genomic DNA
Nuclease free H2O
to 25 µl
to 50µl
 
 
  1. Gently mix the reaction and spin down in microcentrifuge.If the thermocycler does not have a heated cover, add one drop of mineral oil to the reaction tube to prevent evaporation
 
Initial Denaturation
94-95oC
1-5 min
25-40 cycles
94-95oC
45-70oC
72oC
15-30 sec
10-30 sec
1 min per 1000 base pairs
Final extension
72oC
5 min
Final Soak
4oC

 

 

 

 

 

 

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