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missing 500 bp band w/ lambda hind3

daj (David Johnston) daj at nhm.ac.uk
Thu Jul 7 09:25:57 EST 1994


On Thu, 7 Jul 1994 13:03:27 GMT,
  BRIAN PETER MERRY 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.

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.
DAJ

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)



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