qPCR - running taqman for endogenous control and SYBR for the gene - (Jul/16/2008 )
Hi,
Is it possible to mix chemistry when measuring the expression of a gene. Can I use an 18s Taqman probe for the endogenous control and SYBR green for the gene of interest? If this is possible can I run both reactions in a single plate/ single run. (we use the AB 7000 system).
Thanks.
widu
-widu-
QUOTE (widu @ Jul 17 2008, 02:16 AM)
Hi,
Is it possible to mix chemistry when measuring the expression of a gene. Can I use an 18s Taqman probe for the endogenous control and SYBR green for the gene of interest? If this is possible can I run both reactions in a single plate/ single run. (we use the AB 7000 system).
Thanks.
widu
Is it possible to mix chemistry when measuring the expression of a gene. Can I use an 18s Taqman probe for the endogenous control and SYBR green for the gene of interest? If this is possible can I run both reactions in a single plate/ single run. (we use the AB 7000 system).
Thanks.
widu
I am not sure what do you mean by "mixed" chemistry. If you referr to a kind of - multiplexing, then - no. If you have to separate set of reaction, and would like to run them in parallel, in the same plate, then - yes, it is possible.
-Pallas-
QUOTE (Pallas @ Jul 17 2008, 01:11 AM)
QUOTE (widu @ Jul 17 2008, 02:16 AM)
Hi,
Is it possible to mix chemistry when measuring the expression of a gene. Can I use an 18s Taqman probe for the endogenous control and SYBR green for the gene of interest? If this is possible can I run both reactions in a single plate/ single run. (we use the AB 7000 system).
Thanks.
widu
Is it possible to mix chemistry when measuring the expression of a gene. Can I use an 18s Taqman probe for the endogenous control and SYBR green for the gene of interest? If this is possible can I run both reactions in a single plate/ single run. (we use the AB 7000 system).
Thanks.
widu
I am not sure what do you mean by "mixed" chemistry. If you referr to a kind of - multiplexing, then - no. If you have to separate set of reaction, and would like to run them in parallel, in the same plate, then - yes, it is possible.
thanks pallas for the reply... well i am a newbie for qPCR. I am supposed to quantify the expression of a gene. The issue is that the 18s probes (my endogenous control) is Taqman while the gene probes are SYBR green. I'm worried that my data would not be reliable as I am using two chemistries, one for the endogenous control and another for the gene of interest?
-widu-