DNA extraction from polysaccharide-laden plant tissue

Chi-kuang Wen ckwen at expert.cc.purdue.edu
Mon Dec 9 23:36:45 EST 1996


: Steve Pechous <swp108 at psu.edu> wrote:
: Hi everyone,
: 	     I have been trying to isolate DNA from a climbing vine
: (Virginia creeper). This tissue (young leaf) contains very high levels 
: of 
: polysaccharides. After CTAB/chloroform extraction, I use
: 5M NaCL to remove sugars. I have has some success (one prep. worked fine 
: as a template 
i

Hi, Steve:

You said that the CTAB/CHCl3 ext did not work well.  Did you use IPA to ppt
the nucleic acids or use the so-called low-salt buffer?  My experience is
that if you use IPA to ppt the nucleic acids, you also get the polysaccharides.
Low-salt buffer gives cleaner DNA/RNA.

There is a paper, I could not remember where I got it, mentioned that at
high salt conditions, the polysaccharides may bind to the cellulose and
you may pass your DNA through a cellulose column to get clean DNA.

hope this help,

Chi-Kuang
 





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