what's the best probe to quantitate RNA in Northern blot????
helgew at ljcrf.edu
Thu Jan 6 14:32:44 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
>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.
>mman at molbio.cbs.umn.edu
I had the same experience. When comparing actin and GAPDH it got even more
confusing.... right now I am using L32-4A as a standard. The message for
this ribosomal protein seems to be fairly unregulated (at least in the
cells and tissues I am using). You might want to look into Dudov and Perry,
Cell 37: 457 (1984)
as for the rRNA probe, i have also seen people using these, but that seems
to me as good as an EtBr stained gel... :)
my OD260 = 0.02 worth
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