erase a base

KRUEGER, CHARLES M krueger at zeus.tamu.edu
Tue Mar 8 20:06:00 EST 1994


In article <hotan-080394162111 at pc10.hrfs.uiuc.edu>, hotan@@uxa.cso.uiuc.edu (Douglas T.) writes...
> 
> 
> Does anyone have any comments on the erase a base system? 
Greetings,
	Three years ago I had a large cloned fragment in a
plasmid (pK19, pUC19, or pKK388-1; I don't remember
which anymore) that had been identified from a library
via a 32P labeled oligo probe.  I wanted to start
sequencing the clone near the hybridization location
of the probe.  I bought the erase-a-base kit from 
Promega.  I followed the kit instructions .  I think
there was one error in the manual that was easily
cleared up by a phone call to Promega (hopefully the
manual has been corrected by now).  The kit in my hands
performed exactly as advertised.  The nested deletions
resulted in a beautiful desending latter of plasmid
sizes corresponding to the length of digestion.  I used
my labeled probe on a Southern blot made from the gel.
I could determine the plasmid sample at which I had
already digested off the area of interest due to the 
lack of signal from the probe.  I backed up to the next
larger plasmid size and began sequencing.  It worked
great!  I have not had a good reason (shucks) to try it
again, so maybe I was just lucky.
				Charles Krueger
				Texas A&M Univ.
				College Station, Texas



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