DNA size markers

Robin Beech robin_beech at maclan.mcgill.ca
Wed Nov 13 11:02:28 EST 1996


In article <569l0h$2prt at sat.ipp-garching.mpg.de>, xie at made.biochem.mpg.de
(Youhua Xie) wrote:

>: >>molecular weight size marker for our RAPD gels. The cost is mounting as 
>: >>we turn more and more to RAPD technology. Can anyone suggest an easily 
>: >>made DNA/restriction enzyme combination that could substitute for the 
>: >>commercial marker set? I would prefer a sequenced plasmid as plasmid 

We have been using pGEM5z(f+) digested with HpaII which gives a nice range
of fragments from 700bp and down. If you want larger bands you could use an
EcoRV digest for 3001 bp and a PvuI digest for a 2000 and 1000 bp fragment.
We digest with each separately, phenol extract to kill the enzymes then mix
the digests together. This give a marker with nicely spaced bands from 3000
bp down.

Robin Beech
Institute of Parasitology
McGill University



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