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missing 500 bp band w/ lambda hind3

futers at biovax.leeds.ac.uk futers at biovax.leeds.ac.uk
Thu Jul 7 10:59:30 EST 1994

In article <2vh39l$k3r at mserv1.dl.ac.uk>, (David Johnston) daj <daj at nhm.ac.uk> writes:
>On Thu, 7 Jul 1994 13:03:27 GMT,
>>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.
>If my memory serves me correctly, the 564 base fragment and the 23130 base 
>fragment contain the 2 cos sites/sticky ends of the phage DNA and anneal 
>together. Heat marker to 65C for 5 mins and cool rapidly on ice before 
>loading to dissassociate the 2 fragments.
>David A. Johnston
>Dept of Zoology, The Natural History Museum, Cromwell Road,
>South Kensington, London SW7 5DB. England
>(tel 071 9389297, fax 071 9388754, email daj at nhm.ac.uk)

It is the 23130 and the 4361 bands that have the cos sites.  The 564 band is
weak due to being small.  If you need to have a size reference around 500, I
would suggest using a different marker such as the 100bp ladder.

               Simon Futers (futers at biovax.leeds.ac.uk)

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