RNA Pol II CHIP protocol help - suggest protocol (Oct/18/2010 )
I am trying to standardize RNA Pol II chip in the lab. I have tried 2 different protocols, but when i check the enrichments after pcr I see similar intensity bands in the no antibody control and the antibody positive sample, also i sonicate in 0.1% sds lysis buffer and in this condition even after 80 pulses i cant get rid of the >800bp fragments although 60% of the DNA is fragment size is from 500 to 100 bp.
can anyone suggest me a protocol that is standardized for RNA PolII chip. and if i can use a different lysis buffer for sonication?
can anyone share a RNA pol II chip protocol i need it urgently!!
same here.am trying to standardize the ChIP protocol using Ez-ChIP's method.
any help would be appreciated about RNA Pol II's standardization.