what's the best probe to quantitate RNA in Northern blot????

Michael Man mman at molbio.cbs.umn.edu
Thu Jan 6 12:16:18 EST 1994


I am studying expression of a mammalian gene at transcription level, under
different nutritional condition.  I used actin as a probe to quantitate RNA, to ensure equal loading in my Northern.  But I didn't have good luck with actin
probe.  So I switched to GAPDH probe.  My bands looked nicer, but since GAPDH
is also under hormonal control, I can't use it as a quanitative marker either.
Does anyone have experience with ribosomal RNA (15S?) probe in this regard?
I will appreciate if you can share your experience here.

Why not just use 10 or 20 ug of total RNA in each lane?  Maybe you ask.
Yaaaaaa.  I like to hear your logic too.

michael
mman at molbio.cbs.umn.edu



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