new to qPCR - using sybr green (Sep/23/2008 )
I'm new to qPCR. I will be cheking the expression of several genes in my cell lines. I've just ordered my primers. WOuld it be cheaper to check out these primers on regular RT-PCR followed by running gel and then to attempt the sybr green. Can the RT-PCR annealing the,p, denaturation temp etc be used for qPCR as well using sybr green?
Just go directly run sybr green real-time PCR to optimize primer concentration, annealing temperature.
It helps if you can first run a regular PCR with the new primers to see if they give you the specific bands.
U can also look at a dissociation curve after the SYBR run. You should only have 1 peak if you have 1 product.
Multiple peaks means you have multiple products, which will skew your results.