32P-probe by PCR?
Markus.Piotrowski at rz.ruhr-uni-bochum.de
Sat Jul 6 16:44:48 EST 1996
although this should be a FAQ, I was not able to find the older
discussions about this item.
Okay, I want to prepare a 32P-probe by use of PCR. I did it in the past
by simply avoiding the cold dATP and adding 2.5 ul of 32P-dATP (3000
Ci/mmol, 10 uCi/ul) in an total volume of 50 ul. The probes I got had
normally activities of about 20-30 million cpm. Now I want to do
Southerns and people told me that this probes are not hot enough. I know
I should add cold dATP, but I don't know how. Has anyone a PCR-protocoll
that results in higher labelled probes?
I know that I can do ONP-labeling, but I think PCR should work as well.
Many thanks in advance, Markus
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