algal DNA purification
dunahayt at TCPLINK.NREL.GOV
dunahayt at TCPLINK.NREL.GOV
Thu Apr 14 21:50:33 EST 1994
Hi,
We need to isolate total DNA from small volumes of large numbers of
microalgal cultures for analysis by Southern blotting and PCR. We've been
using a miniprep method (obtained from the Chlamydomonas Gopher) using
phenol/chloroform extraction, which gives decent yields, but digestion with
restriction enzymes is usually poor. We think this is due to carbohydrate
contamination. CTAB precipitation helps the digestion problem, but for
large numbers of samples, CTAB is a nuisance, and yield is very low. Does
anyone know of any methods, or commercial spin columns, or anything, that
allows easy clean up (carbohydrate removal) of genomic DNA? I'm going to
try the Puregene system from Gentra Systems (DNA isolation without
phenol/chloroform), but I'm afraid that this might not address the
carbohydrate problem.
Any info would be appreciated.
Terri Dunahay
National Renewable Energy Lab
dunahayt at tcplink.nrel.gov
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