Dear Marco,
you are correct, the labeling step is not in protocol even though it was in the version that I had submitted. Sorry for that. The missing part is:
3) Labelling reaction (20 µl)
Dephosphorylated RNA 1 µg
T4 Polynucleotide Kinase buffer 10X 2 µl
RNase inhibitor (40 U/µl) 0.25 µl
?32P ATP (10 µCi/µl) 5 µl?
T4 Polynucleotide Kinase (10U/µl) 1.25 µl
H2O to 20 µl final volume
a. incubate 2 hours at 37°C
b. Precipitation as in step (2) and take up the pellet in desired volume
I will ask to correct this on the web site.
Harald
2013-10-24 02:54:45