I have a set of primers that works well in a regular PCR (using QIAGEN multiplex master mix) but is not compatible with the qPCR reagents. Redesigning the primers is not an option, so I'm wondering if I could mix in some of my regular PCR reagents with the SYBR green reagents and try out a qPCR?
Hi,
I guess, it should work!
I quite often use side-qPCR reaction to define the concentration of the NGS libraries in the PCR mix (PCR master mix + DNA + 2x Primers). After a couple of initial cycles I take 5ul of this mix and add to the SYBR qPCR master mix and run qPCR to define the amount of cycles required to get saturation. Works fine for me.
Best,
Michail
Hi,
I would not mix different PCR mixes but it may work. However, you can buy SybrGreen or similar dyes seperately and add it to your Qiagen mix.
Good luck,
Norman
Hi Ishani,
what is the reason for non compatiblity of the primers with Sybr chemistry? In general PCR Reagents are developed for particular applications so this might have side effects one cannot forsee right now. Let me know if you amy discuss in more detail
Sven
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