I used GENET BIO (ExPrime Tag Premix (2x)) for my PCR rxn.
It contains: DNA Polymerase 1 Unit/10 microliter, 20 mM Tris-HCL, 80 mM KCl, 4mM Mgcl2, enzyme stabilizer, sediment loading dye, pH- 9.0 and 0.5 mM dATP,dCTP, dGTP, dTTP.
1). After My PCR rxn was completed , what is the pH ?
2). Can I use PCR product as such for my restriction digestion?
3). If the PCR solution interfere or not with my digest?
5). the PCR solution (pH 9.0) it will affect restriction enzyme activity or not?
6). Is there any special product which is available in our market?
Hey Senthil, In our lab we have used different PCR mixes and we directly use them for restriction digestion for ARDRA and cloning. Till today we have not experienced any problem. You can directly use your PCR product. Good luck.
Naveen without elution it is possible that PCR rxn mix contain 20 mM Tris-HCL, 80 mM KCl... it will affect my product r not...
Tris HCL and KCl is present in almost all Taq buffers. From my experience, I don't think it will be a problem....
We have used different types of PCR mixtures and never had a problem. What you can simply do is that compare the results of digestion on two sets of identical samples say 10 each and perform two types of PCRs, one by using this mixture and the other by preparing your own. Compare the results to see any difference, which I expect wont be.
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