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missing 500 bp band w/ lambda hind3

kramerse at wmvx.lvs.dupont.com kramerse at wmvx.lvs.dupont.com
Thu Jul 7 11:02:19 EST 1994

In article <002846m.31.2E1BFD1F at axe.acadiau.ca>, 002846m at axe.acadiau.ca (BRIAN PETER MERRY) writes:
>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
I believe you need more mass.  If you have 500ng of total mass in the lane
you will have about 6 ng of mass in the 564bp band.  I just looked it up
in one of my methods books, it says "20 ng per single band is readily
detectable".  Your best bet is to call the supplier of the marker and
see what they recommend.  Hope this helps.
Susan Kramer

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