Label during RT for Expression Profiling (reverse Northerns) ?

Fredrik Kamme fkhero at biogen.wblab.lu.se
Thu Apr 17 09:49:50 EST 1997


Hi,
I tried both, labelling the cDNA during synthesis and labelling
afterwards using random priming. Both seem to work. You can get a lot of
label into your probe with RTase, so I prefer this as it is easier.  I
used a modification of the protocol published by Erlander et al. in Nucl.
Acids Res, vol 25, pp913-914, 1997. I used 15µg of total RNA, 800µM
dNTP-dCTP and 7.5µl of dCTP (3000Ci/mmol) in a 25µl reax. and got a total
of approx. 200x10^6 cpm.

Freddy



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