oligo cloning

Caroline Szymeczek-Seay css at med.unc.edu
Mon Feb 7 14:25:41 EST 2000


I'm cloning (or trying to clone) a double stranded blunt ended oligo (a
47mer) into pGL3 enhancer vector from Promega.  Vector is cut with SmaI
and CIPed.  Oligos are kinased.  

My question:  is there a rule of thumb for amount of oligos to add to
vector?  I'm trying a 10-fold molar excess, a 5-fold molar excess and an
equimolar amt of oligo relative to vector. I'm getting no positives. 
This is a little hairy also because there's no blue/white screening for
this vector.  


caroline




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