Dear all,
I should extract DNA from some cancer cell lines and perform PCR for its. I did it by Qiagen kit. From among all, PCR of DNA from Mia-paca-2 cell line (cell line of Pancreatic cancer) didn't give me single band ( I tested it with different primers for housekeeping genes and some others). Although, extracted DNA runs on the gel and quality of it is OK.
can any one help me about this problem?
Is there special extraction method for this cell line? or do this cell has special inhibitor material in extract?
Post a picture of your PCR that will make it easier to answer your question.
Dear Marziea,
It sounds like you got PCR amplification from DNA extracted from other cell lines with the same primers and that you used a number of differnt primers, so really, it sounds like there is a problem with this particular prep.
I recommend that you repeat the DNA extraction from this cell line and re-do your PCR with the new extracted DNA. Sometimes it can be a simple mixup during prep and it really isn't worth it to spend weeks trouble shooting when a simple repeat may result in success.
If that still doesn't work, let us know and we can attempt to help with trouble shooting...
Dear Petra,
thank you for your following
I repeated it several times, but PCR of this cell didn't give me any bonds, exceptionally, I can get a sharp band with GAPDH primer from cDNA synthesis kit of Fermentas whereas other cells such as Pa-tu-8902 and ... give sharp bonds with all primers, this means that all primers are OK. I'm really confused
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