Primers and Taqman Probes mixture question - (Jul/26/2009 )
So at my place of work, someone has questioned the integrity of a primer probe cocktail (fwd, rev primers & taq man probe). I've been doing PCR, RT-PCR and Real Time PCR for just about 6 years now and when possible, I've mixed the fwd and rev primers into a singular cocktail. It was easy to maintain and most importantly, it worked.
I usually make a cocktail (75 ul) that lasts me about a week without any problem either. I store the primer/probe cocktail at 4 degrees C during the time I use it. The primers and probes are entirely specific to their target sites and non complementary. So what could possibly go wrong?
-Joe
-jfrezzo-
jfrezzo on Jul 27 2009, 12:39 AM said:
So at my place of work, someone has questioned the integrity of a primer probe cocktail (fwd, rev primers & taq man probe). I've been doing PCR, RT-PCR and Real Time PCR for just about 6 years now and when possible, I've mixed the fwd and rev primers into a singular cocktail. It was easy to maintain and most importantly, it worked.
I usually make a cocktail (75 ul) that lasts me about a week without any problem either. I store the primer/probe cocktail at 4 degrees C during the time I use it. The primers and probes are entirely specific to their target sites and non complementary. So what could possibly go wrong?
-Joe
I usually make a cocktail (75 ul) that lasts me about a week without any problem either. I store the primer/probe cocktail at 4 degrees C during the time I use it. The primers and probes are entirely specific to their target sites and non complementary. So what could possibly go wrong?
-Joe
i don't see any problem since you have so much experience and you cocktail seems to be so robust. If there's no problem, why find one? If you are more experienced than your colleague explain all the precautions in taqman PCR, and if the results are fine, what's the problem???
chris
-chrisbelle-