DNA from cartilage?
wolffie at u.washington.edu
wolffie at u.washington.edu
Fri Jul 2 18:46:31 EST 1993
I'm extracting DNA from cartilage for Southern blotting and PCR.
Lots of glycoprotein or other such white glop coprecipitates with the DNA.
I don't know how much of a problem this is going to be, but I'd like to get
rid of the stuff if I can.
Any ideas?
The cartilage is minced, digested with proteinase K in 1% SDS-STE (sometimes
predigested with pronase or collagenase). Then we add 1/3 vol of saturated
NaCl and extract this with Chloroform/isoamyl alcohol and precipitate with
isopropanol (we don't use phenol anymore). The flocculent white precipitate
doesn't look like DNA and is much larger than one would expect; although it
does contain DNA, I expect this stuff is mostly digested glycoprotein
matrix.
Thanks,
John Wolff,
University of Washington
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