Any good: Invitrogen Subtractor Kit? (biotinylation-based)

Dr. Hooper F. timt at itsa.ucsf.edu
Fri May 9 16:51:52 EST 1997


Trying to do a subtractive cDNA library, I am soliciting advice on what
commercial kit to choose.

Invitrogen has a Subtractor kit that (1) the uninduced RNA is biotinylated
first and then hybridize to the induced cDNA pool, and (2) the hybrids of
common sequences, now with biotin, bind to streptoavidin and then removed
by phenol extraction. The induced cDNAs, being unique to induction, will
not hybridize to the uninduced RNAs and thus stay in the sup during phenol
extraction.

Anyone has good experience with the kit?

Wouldn't phenol extraction denature streptoavidin and break the
biotin-streptoavidin complex and thus leave behind the commoners?

What are the weakest features of the kit?

Many thanks for your attention/comments!  Tim



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