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missing 500 bp band w/ lambda hind3

Dan Rohwer-Nutter DAN at biosci.bsd.uchicago.edu
Tue Jul 12 15:40:34 EST 1994


In <002846m.31.2E1BFD1F at axe.acadiau.ca> 002846m at axe.acadiau.ca writes:

> 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.
> 
> Brian Merry

Have you tried heating the markers?  I know that two of the upper 
fragments will partially re-anneal and run as one band if the digest isn't 
heated to 56C for about five minutes.  Maybe this is also happening to the 
smaller fragment?

Good Luck!  Dan.




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