We have used Premix Ex Taq (Probe qPCR) Master Mix. Takara Bio Premix Ex Taq (Probe qPCR) for awhile, but recently found that it didn't give satisfactory results, so we switched to TaqMan™ Multiplex Master Mix(Thermo Fisher) without success. Strangely, the latter enzyme gave amplifications about 100 times more than the former enzyme. We suspect that the latter enzyme has non specific amplification problem. Could you recommend a good Probe qPCR Master Mix?
Jia-Wang: The key to a successful qPCR is the amount of high quality SYBRE green contained in the master mix. Thus, it can supply enough dye for high copy cDNAs pushing the CT value well below 12 in a linear fashion. I have compared a few master mixes from various vendors, and found the one from Roche met our needs. (Disclaimer: no compensation of any kind received from Roche Corporation). Good luck.
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