Hello
I have been extracting DNA from PBMCs and sorted CD4+ cells for PCR, unfortunately the A260/280 readings are between 2.6-3.3 and the A260/230 are of similar values.
Can anyone explain why this is and what I can do to make the ratios nearer 2?
I have ruled out the freezing media, I was wondering if it could be ficoll carry over? Or a high amount of RNA ( I don't have an RNase step)? I'm lost!
The samples do not perform well in the PCR, whereas extracted buffy coats with A260/280 of 1.9-2 amd A260/230 of the same values work beautifully.
Thank you!!
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