PCR of semen samples
ANDREI.POPOV at afrc.ac.uk
Tue Dec 6 13:49:18 EST 1994
I use high-molecular weight DNA from human sperm cells;
though I prepare my DNA in agarose blocks, the same
protocol should work in solution.
Wash cells twice in PBS; resuspend at ~25x10-6 cells/ml
in 1% Lauroylsarcosinate, 10-20 mM 2-ME, 25 mM EDTA,
1 mg/ml Proteinase K (Sigma P4914). Incubate o/n at 56C.
PMSF-treat, extract, ppt....
There are 2 key points: 2-ME and high concentration
of active Pr.K.
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