The best CHEF electrophoresis conditions??

Nelson Salazar N.SALAZAR at lshtm.ac.uk
Sun Jun 13 15:44:25 EST 1993


Hi Netters,
            I am interested in separating the Entamoeba histolytica
"chromosomes" by CHEF electrophoresis and I would appreciate it very
much if someone knows how can I "predict" the best CHEF conditions
to get a nice resolution of these "chromosomes".

1) Voltage?
2) Running time ?
3) Ramping pulses (Initial and final times??)

The E. histolytica "chromosomes" are about 0.3 to over 3 MegaBases in
size. Another thing that is worrying me a lot, is the fact that I am
getting a smear of DNA and therefore this makes very difficult to see
the bands when stained with EtBr if there are any at all !!

Any suggestions are welcome. Thanks very much in advance.

                        N. Salazar







**********************************************************************
Nelson Salazar                      *  E-mail: n.salazar at lshtm.ac.uk
Dept. of Medical Parasitology       *    or    nsalazar at mrc-crc.ac.uk
Lon. Sch. Hyg. & Trop. Med.         *  Phone : 071 927 2468 (Office)
Keppel St. London WC1E 7HT, UK      *  Fax   : 071 636 8739
**********************************************************************



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