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missing 500 bp band w/ lambda hind3 (fwd)

Jason Schneider schneid at rs6.tcs.tulane.edu
Thu Jul 14 14:45:26 EST 1994

Robert Preston (rapr at MED.PITT.EDU) wrote:
: > 
: > Hi,
: > We need some help.  We're running 1% and 2% gels in our lab using Lambda hin3 
: > as a molecular weight marker.  We load 500 ng of lambda in our marker lane and 
: > we don't get resolution of the 500 bp band when prestaining the gel with etbr.
: > Do we need more DNA in the marker lane?  More etbr (we now use 2ul/50ml of 
: > gel, this used to be enough).  Any comments/suggestions will be appreciated.
: > 
Are you heating the marker before you load it? Lambda has cos ends that really
like to link up. If one of the bands on your lane is unusually bright, that
500 bp fragment may mixed up in there. Heating at 65^C for 10 minutes should
do the trick. If that is indeed your problem.
Jason S. Schneider
Tulane University
schneid at mailhost.tcs.tulane.edu

"With imagination, I'll get there..."
           --Harry Connick, Jr.

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