reference on PCR using degenerate primers - Help me find the reference (Nov/19/2002 )
Hi, I'm writing a thesis but I couldn't find a reference on PCR using degenerate primers. No product was observed from first PCR but when I used that solution from first PCR as template in nested PCR, I got a band. I need a reference to write that the product from PCR using degenerate primers can be a smear or a product which couldn't be detected with ethidium bromide staining. Can you suggest me websites or textbook. Thank you.
In nested PCR, it is a normal phenomena. That's why we do nested PCR.
In first PCR, result will be a smear if we want to get a band by increase the number of cycles or quantity of template, because first PCR is moderately specific. We do Second PCR to increase specificity.
You may search some references about nested PCR.