Erase a Base

[Sheryl White]

rick lynch (vaxkiller.agi.org!rlynch) wrote:

: I need to generate sets of nested deletions.  I have been looking into a kit  
: sold by Promega called "Erase-a-Base"  The company claims it is one of..  
: wonderful nested deletion kit on the market, but if I had a nickel for every  
: time I heard that I wouldn't be working.:-)
: Anyone tryed it?  If so, could you please let me know what you think of it.

: Thanks

: Rick Lynch
: Agouron Institute
: rlynch at vaxkiller.agi.org

I've successfully used the Erase-A-Base kit.  At the time, it worked 
fairly well for making deletion constructs of a fairly large cDNA.  
However, I found it a bit tedious: after the exonuclease digestion and 
subsequent ligation into your vector you still need to check out quite a 
few of the colonies/timepoint to find one the right size (the timepoints 
sometimes give a very wide range of deletions, from 0 to 1 kb, on 
occasion). I find it's alot easier and faster to generate deletions using 
PCR and nested primers, I have yet to find a transformant which wasn't 
exactly the size it was supposed to be.  Hope this was helpful...

----------------------- Sheryl L White  -------------------------
    Dept of Mol Physiology, Given Med, UVM, Burlington, VT 05405
       Voice: (802) 656-4433  E-Mail: slw at salus.med.uvm.edu
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