problem with detection limit on electrophoresis mobility shift gel
AEVDOKIMOZ at cinci.rr.com
Thu Feb 28 22:55:19 EST 2002
1) rnases ? could it be that your rna is digested before it even gets to be
anywhere ? rnases are a MAJOR pain.
2) chaotropes may help you.
"shchan" <chanhongsiu at yahoo.com> wrote in message
news:a5cr97$bqs$1 at justice.csc.cuhk.edu.hk...
> I am working on gel shift assays for RNA-protein binding. Many people
> published their gel shift using just 0.02 nM of 32P-labelled RNA but I can
> see nothing at this concentration even I used 32P of ~3000Ci/mmol
> specificity activity. Anyone can help?
> Plus, I am transcribing some RNA of very high secondary from oligo DNAs.
> melting temperatures of those 30-mers are ~90 deg/C. I found it difficult
> make good yield on them. Anyone can help?
> SH Chan
> Department of Biochemistry
> The Chinese Unvversity of Hong Kong
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