Degraded DNA Cleanup?

Dr. Duncan Clark duncan at genesys.demon.co.uk
Thu Jan 23 04:12:52 EST 1997


In article <5c01fq$jfa at news.doit.wisc.edu>, Dima Klenchin <klenchin at facs
taff.REMOVE_TO_REPLY.wisc.edu> writes
>In article <32E397D7.60C8 at GENVEC.COM>, CANTOLUPO at GENVEC.COM (Lou Cantolupo) 
>wrote:
>->Anyone have a good protocol to clean up degraded DNA?  My fragment is
>->about 35Kb (whole adenovirus DNA), VERY PRECIOUS, and is marred by a lot
>->of degraded DNA.  I'd like to get rid of all the junk.  I've
>->reprecipitated and that didn't help much.  Any suggestions?  Thanx in
>->advance!!
>
>I would try cesium gradient. 

I doubt AD2 will separate from degraded DNA on CsCl as they will both be
linear. If your degraded fragments are smaller then I would gel purify
your large fragment. If you have a lot of sample, sucrose gradient
should size select OK. If you only have trace amounts then maybe long
PCR of the whole lot or spli it into 17.5kb or 12kb chunks. 

Duncan
--------------------------------------------------------------------------------
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Dr. Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.com
http://www.genesys.demon.co.uk



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