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Pulsed field GE of large circular DN

Ronan.OKennedy at UCG.IE Ronan.OKennedy at UCG.IE
Sun Jul 24 14:50:08 EST 1994


I have had some replies concerning my query of using PFGE to seperate large
circular (bacterial ) chromosome. What i would like be able to is where a 
culture contains a mixture of bugs (Eg E.coli Coryne. glutacium, Bacillus, 
Vibrio) that PFGE would sererate out four circular chromosomes and mega plasmid
if present. The info I have suggests that reduction in field strength and use
of pulses ramped from 5sec to 240sec increases the mobility of circular DNA 
Although the DNA from the cultures above cultures enter the gel there is little
if not no difference in their mobilities. One suggestion that I have just noticed is the use of gamma irradiation to introduce double strand breaks into large 
DNA molecules. I still have to try this 
Again any advise would be appriciated
Many thanks
Ronan O'Kennedy
Dept of Microbiology
University College Galway.



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