rRNA visualization in formaldehyde gels
rh at mblab.gla.ac.uk
Mon Oct 23 18:52:32 EST 2000
In article <email@example.com>,
mlsulliv at facstaff.wisc.edu ("Michael L. Sullivan") wrote:
> >I can not see any rRNA on the formaldehyde gel.
> >The protocol is the one in Current Protocols : agarose gel, formaldehyde and
> >MOPS. The loading buffer is sterile filtrated and then DEPC treated and
Why not use a test gel with 1% agarose and denature the RNA for 10 mins @
60C in 3:1 formaldehyde/formamide then run for 1 hour @ 75V.
You should see the bands no problem. PS you could add EtBr to the gell at
the conc below.
Cyber Green is more sensitive. Ask Molecular Probes for freebie. :-)
> Add EtBr to either the sample (as Susan suggested) or to the gel and buffer
> (I use 10 ug per 100 ml). Staining after running I think is less sensitive
> (most people don't have the patience to stain and de-stain, plus diffusion
> might be a problem).
> >Is the detection level in the range 500 ng or can I see like 20 ng?
> Certainly 500 ng would be pretty easily seen. 20 ng is probably too little
> to see. I think there are stains that might be more sensitive than EtBr
> though (Cyber green maybe?), if you really need to see less. Hope this
Bob; Rainy Scotland
Robert Hartley, Centre for Cell Engineering,Joseph Black Building
University of Glasgow, Glasgow G12 8QQ Tel: ++44 (0)141 330 4756,
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