strange cloning results

William Sun william at neuro.usc.edu
Fri Jun 12 12:37:24 EST 1998


Hello all,

I've isolated several clones from a cDNA library with a labeled probe.  When
I analyzed the clones, I found that many of them are in the reversed
orientation (the library was oligo-dT primed and uni-directionally cloned).
Is this just an artifact or can some gene have 2 ORFs in opposite directions?
Another strange clone that I isolated was that if I translate the aa in the
same frame as an already known homologous protein, I don't detect a stop codon -
it translates right through to the poly-A tail.  I sequeced the clone in both
directions so I'm sure it's not a mistake in the sequencing.  Can anyone offer
an explanation? 

-- 
William Sun, Ph.D               
Metabasis Therapeutics, Inc.           Phone: (619) 622-3930
9390 Towne Centre Drive                FAX:   (619) 622-5545
San Diego, CA 92121                    Email: william at alum.mit.edu 



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