Problem with PCR on DNA from paraffin section - (Dec/22/2005 )
I have problems with my PCR. I work with DNA extracted from tumors embebed in paraffin. So, the quality of my DNA is not good. It has a lot of contamination. A month ago, I amplified this DNA and I obtained the product I needed. This week, I repeated the reaction with the same protocol, but I didn´t obtained anything. Therefore, I repeated the reaction a lot of times. Also, I changed the concentration of primers, MgCl2, DNA......., but it didn´t work. I don´t know what the problem could be?? Could anyone help me??
PCR can be a pain in the ass sometimes cant it!?
I had a similar problem recently, I would run the DNA on a gel to ensure that it is not too fragmented and as long as there is still some DNA maybe re-purify it and try again!?
all the best!
Try re-precipitating your DNA, washing and re-suspending in buffer. Ususally works for me when such samples (paraffin embedded) start going wrong