dede, on Sep 2 2009, 11:05 PM, said:
Hi all
Some of my Genomic DNA samples are not giving pcr product.....there are no protein contamination.....OD is 1.8
right now i am using 10ng/ul of dna
i am unable to find out the reason behind this problem.
...even i did reprecipitation of dna but still no result.
I am wondering if i cud change the buffer to dissolve DNA ,right now i m dissolving it in TE buffer
to clarify:
some samples are not yielding any product, but separate independent samples are yielding product when assayed with the same primer pair...or one primer pair yields a product, but another primer pair when used with the same gDNA does not yield a product?
i'll assume the former rather than the latter.
are these diagnostics in that you're looking for variations in alleles (e.g. insertions, deletions, CNV etc.)?
if you're not already doing so, you should always run a control PCR rxn along with your experimental rxn's. the control is simply ensuring that your gDNA is of PCR quality...aside from the OD readings, have you looked at your sample on a gel to ensure it is not degraded?
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