I sent this message yesterday, but it didn't show up at my address so I
think it got lost in E-mail hyperspace. (If it shows up at your address
twice, I apologize).
I'm trying to isolate small amounts of total DNA from a large number of
isolates of an alga for analysis by Southern hybridization and PCR. We've
been using a "miniprep" adapted from a Chlamydomonas protocol we got from
the Chlamy Gopher; basically a SDS/phenol/chloroform extraction. We get
good yields, but the DNA digests very poorly with restriction enzymes,
presumably due to inhibition by carbohydrates. CTAB precipitation will get
rid of the carbohydrates, but the protocol is a pain for numerous small
samples, and the yield is very low. My questions: does anyone know of an
easy protocol or a commerial product (i.e., a spin column) that will get
rid of carbohydrate from a DNA prep? Even better, would be a quick kit or
something that yields digestable DNA without phenol/chloroform extractions.
I spoke to someone about using th Puregene isolation kit (involves a
protein precipitation instead of PCI), but I'm not sure it will help with
Thanks for your help.
National Renewable Energy Lab
e-mail: dunahayt at tcplink.nrel.gov