DNA Extraction Protocols for Campanulaceae

Darren Crayn P8821860 at UNSW.EDU.AU
Wed Feb 19 09:41:49 CST 1997


>Can anyone suggest an improvement to the standard
>Doyle & Doyle (1987) protocol for the removal of
>polysaccharides. The Campanulaceae seem to be
>particularly bedevilled by these and although I
>have successfully extracted and amplified good
>DNA from about two dozen genera, the whole procedure
>is time-consuming and decidedly unpredictable.
>I have tried precipitation with 2.5M NaCl in addition
>to other protocols such as Dellaporta (which
>involves chloroform/phenol extraction) with only
>marginal improvement.I'm still left with a
>gelatinous mass (or mess !).
>
>Bill Eddie

Have you tried using a lithium chloride protocol? I believe there are some
molecular phycologists in our department who have had an inordinate amount
of trouble getiing DNA rather than polysaccharides out of fresh tissue.
>From memory, I think using lithium chloride has been to their benefit. I
know no more about their protocols but I will check up on it and get back
to the list. In the meantime, does anybody else know anything about this?

Cheers
Darren Crayn




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