Megabase DNAs and PFGE?
riggs at utsc.utoronto.ca
Thu Jun 26 13:37:10 EST 2003
We are interested in using pulsed field gel electrophoresis to separate
large fragments (approx. 0.2 to 1Mbp) of Arabidopsis DNA. A look
through the databases reveals that very little has been published on
this. Can anyone provide practical advice on the following questions:
1. How does one prepare megabase-sized DNA? Do standard leaf
protoplast/agarose embedding procedures work well?
2. Does anyone have any experience in using restriction enzymes which
cut infrequently (and preferably not at the whim of methylation?); has
anyone tried homing endonucleases?
3. Any advice for PFGE parameters?
Thanks for your help!
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