1 reply to this topic

[angel0071987]

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

 

Attached Thumbnails

• 

[Astilius]

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).