Hello all,
Hope this mail reaches you all.. I have a couple of
questions about 2D gels.
I have been running 2D gels for quite sometime now. Before I
pose my question, certain details about my protocol:
I use multiphor for focussing, Protean II for SDS
page...large format gels. I use Chaps, Dnase, Rnase in my
sample prep. I have also tried Acetone precipitation. I load
sample by in-gel rehydration. My problem is with streaking.
There are always streaks near the acidic end (mainly) and
near the high molecular weight region. Though I have tried
different sample cleanup methods, I still continue to have
streaks but I have been succesfull in reducing them quite a
bit.
1. Could someone give me the amount of DNase/RNase used in
proportion to the sample?
2. Will sample cup loading help in reducing streaks (in the
sense, the streaks wont be spread throughout the length of
the strip as in the case of sample loading by in-gel
rehydration?)
Please help me with this problem. After all the efforts
in running 2D gels, I would love to have great looking gels!
Babur
This archive was generated by hypermail 2b29 : Wed Nov 21 2001 - 14:16:55 EST