Poly(A)+ selction

frist at ccu.umanitoba.ca frist at ccu.umanitoba.ca
Wed Oct 23 09:41:44 EST 1991


In article <0095080B.B0FCAEA0 at m44.unm.edu> kim at m44.unm.edu writes:
>Dear group:
>
>  I am trying to do Poly(A)+ selection of RNA using either Oligo(dT) cellulose
>or Amersham Messenger Affinity Paper.  I have been having a lot of trouble with
>the procedure, and I need some advice.

... deleted description of method ...

>  When I run the "Poly(A)+" RNA on a gel, however, there is no attenuation of
>the rRNA bands, compared with an equivalent mass of total RNA.  Nor is there an
>enhancement of these bands in the Poly(A)- fraction.  It is as if there is no
>specific binding at all.  My yields, if that is the correct term, are around 1%
>to 3%  of the total RNA applied.
>
>Thanks in advance . . .  Daniel Kim  (KIM at FLOVAX.LANL.GOV)

mRNA complexes with rRNA and these aggregates may need to be broken before
oligo dT chromatography will do any good in enriching for the polyA+
fraction. See

Banile et al. (1976) Analytical Biochem.  72:413-427.
  
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