PFGE agarose plugs

Dr. I. Dunham idunham at crc.ac.uk
Wed Jun 9 11:13:49 EST 1993


I've come across at least four ways of forming plugs for PFGE.

1.  Buy insert moulds from (for us) Pharmacia.  Although this is expensive, you can get
    a lot of them.  The downside is that unless you also get a gel comb from the same 
    company you'll have to make one the right size.

2.  Get your workshop to make you a mould and compatible comb out of perspex.

3.  A neat and cheap trick is to form the plugs in a disposable plastic pasteur pipette.  
    The agarose/cell slurry is sucked into the barrel of the pipette and set on ice. You
    can then release your plugs (we call them worms) by cutting of the narrow part of the
    barrel and squirting the gel/cell mixture in to your tube for lysis.  A disadvantage
    of this method is that although it allows you to do a lot of different samples, all
    your plugs are round and don't fit your gel combs very snugly.

4.  Pour the agarose/cell mixture in to a petri dish to approx 2mm depth, place on ice
    and then cut out squares to make plugs using a razor blade.

Hope this helps 

Ian
idunham at crc.ac.uk 



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