PCR efficiency - (May/30/2007 )

Hi,

My query is on calculating PCR efficiency.

Can you only get a measurement on efficiency from a standard curve??

As, I have 2 genes of interest, and i have serial diluted the cDNA, to see the limit of detectin of these two genes also i am interested in how the efficiency changes as the template copy number decreases.

Hence, its like a standard curve but the fragment is not purified.

Can i get the efficiency from the slope of serial diluted cDNA??????

Thanks

angie

Dear Angie,

You can plot a standard curve using your cDNA and IMHO you should use the exact cDNA to plot it instead of purified fragments, if you're trying to measure the copy number of a cDNA fragment in a cDNA pool

Jahan is right. Just do a serial dilution of your cDNA. The units are arbitrary, so you do not need purified fragments at all.