rRNA visualization in formaldehyde gels

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Mon Oct 23 10:48:08 EST 2000


>Hi.
>
>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
>autoclaved.

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
helps.

Mike


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