DNA quantitation question
Thu Jan 26 13:23:05 EST 1995
This may be quite a basic question, but it has been troubliong me for
I am currently working on a project using PCR to quantitate mutations.
Quantitative PCR has it's own pitfalls, but my question is more
fundamental. I extract DNA from mammalian tissues, using a high salt
protocol and I use a spectrophotometer to calculate my DNA concentration.
I have found that this can be quite unreliable. If your DNA is quite
concentrated, then pippeting it is extremely difficult. Resuspending
your pellet in a large volume helps this, but even then if you repeat it
you often get different values. My question is, is there an established
method for calculating DNA concentation, which is more accurate than
using a spec.
Your thoughts on this would be appreciated
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