DNA extraction from hair - issues on nanodrop - (Sep/06/2016 )
Hi
I have been trying (unsiccesfully) to extract DNA from horse hair samples.
I have tried the Qiagen user developed method, and have had rubbish levels of DNA back (around 2ng/ul). I have recently tried another method using proteinaseK, Tween20, NP40, promega taq buffer and MgCl2 extracted for 60C 45mins, then 95C for 15mins.
I have got the following curve on the nanodrop... really high peak at 230nm and a shift in the DNA peak from 260-280
would cleaning up the DNA work?
Any advice or suggestions greatly appreciated.
Thanks
The peak at 230nm is likely to be not from your DNA (phenol and carbohydrates typically absorb at this wavelength, Trizol gives a response at 230nm and 270nm).
You say 2ng/ul is "rubbish" but neglect to tell us how much DNA should be extracted and what the elution volume is.
You might want to do a 20mg/ml proteinase K, 1M DTT digest and then spin the extract through a suitably spinfilter to concentrate the extract. But put in the amount of intact hair bulbs to give you the amount of DNA you need (whatever that is).