Poly(A)+ isolation tricks

Jim Hill hill at nana.slu.se
Thu Mar 12 09:14:43 EST 1998


> 
> At 14:58 3/11/98 +1000, Wendy Ingram wrote:
> >Hi everyone,
> >
> >I'm about to embark on some Poly(A)+ isolation using oligo(dT)
> >cellulose.  I have protocols etc, but would really like to find out some
> >of the finer details.  Unfortunately I have to do it the old fashioned
> >way (unless someone can suggest a very economical kit :-).
> >
> >- in particular I'd like to know the tricks for scaling things down for
> >small amounts of total RNA (eg how much oligo(dT) per ug of total RNA is
> >needed).  My samples are scarce so ideally I would like to put in about
> >200ug of total RNA and get good enough recovery for a lane on a
> >northern.

Hi Wendy,

I think you may find the Smart cDNA synthesis system from Clontech to 
be of use (no affiliation). You can read the blurb on their website, 
but effectively it allows you to amplify full length cDNAs from small 
amounts of starting total RNA. I have had considerable success with 
it, so if you don't specifically need polyA-RNA and can make do with 
CDNA then its worth a try.

Jim



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