formaldehyde gel for RNA
goa at aber.ac.uk
Thu Jun 20 06:31:10 EST 1996
In article <4q51m2$5cm at nuscc.nus.sg>, zoowyw at leonis.nus.sg says...
>I'd tried to run my RNA sample on formaldehyde agarose as protocol on
>Molecular cloning by Sambrook several times, but I get smear bands for
>samples and marker and failed to get two distinct band for 28S and 18S as
>expected. I'd eliminated the possibility of RNase contamination. Is these
>caused by quality of formaldehyde or pH? Can I have references for these
>Thanks in advance.
If you follow the method and it doesn't work then either you've done it
wrong or else your RNA is crap. Points to note are:
1 Make sure that your formaldehyde is ok and doesn't contain too much
2 Denature the RNA in the formamide/ formaldehye loading buffer for
15 mins at 65 C and then snap on ice.
3 I tend to run low formaldehyde gels now at about 0.6 M.
4 You can put etidium in the loading buffer of stain the gel later
in 0.5 ug/ ml ethidium in water or acetate. I generally transfer and stain
the blot with methylene blue.
Good luck, Gordon.
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