PCR variability with DNA extraction method

P R Orange P.Orange at shef.ac.uk
Mon Aug 5 09:42:20 EST 1996


Hello, I was wondering if any one else has experienced this problem and 
if so, what they did to solve it.

I am using a PCR-restriction digestion assay to genotype some blood 
samples, and I find that one of the polymorphisms is completely 
undetectable when I use the "Nucleon" kit from Scotlab to extract the 
DNA, but when I take the same blood sample, boil for 15 mins, spin out 
the crud and use the supernatant, there is no problem in detecting this 
polymorphism.

I've done extensive experiments to ensure that it is not a variability 
in the PCR or digestion steps, and IT IS NOT,

Please can anyone help me??

Thanks in advance,

Paul Orange (P.Orange at sheffield.ac.uk)



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