Favorite DNA isolation protocol

Gregory Meyer GMEYER at BIO.TAMU.EDU
Wed Nov 8 09:08:25 EST 1995


Our lab works with Cryphonectria parasitica, i.e. chestnut blight, I have
found in my hands that I regularly get 1-1.5mg/ml (I consider this good for
this fungus) using liquid grow up, filtration of the mycelia through
miracloth followed by -70C and an overnight lyophilization.  I use mortar
and pestle without liquid nitrogen (in a hood!  Secondary metabolites are
everywhere) and then do hot alkali lysis followed by phenol:isoamyl
alcohol/chloroform extraction once, isopropanol ppt/centrifuge, resuspend in
TE, RNAase incubate, chloroform:phenol extract 2X, and then once with
chloroform only, ethanol-salt ppt the DNA/centrifuge, wash and resuspend in
TE.  If you would like all the specifics, this may be close to identical to
what you are presently doing and so of no use, just let me know.
Greg Meyer
Biology Department
Texas A&M
E-mail=gmeyer at bio.tamu.edu
























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