DNA extractin from gel HELP

Michael Morales mmorales at acpub.duke.edu
Tue May 2 11:42:48 EST 1995


beavis (beavis at butthead.utah.edu) wrote:
: I've been trying for awhile now to extract a 30Kb piece of DNA from a
: pulse field gel with no success.  Initally I used B-agarase method
: following the protocol that came with the enzyme.  This did not work so I
: switched to Qiagens Quiaex11 kit and still with no success.  It is a 1%
: low melting point agarose gel and we use TBE as a buffer.  The gel used to
: test for presence of DNA  in the extract never shows a band.  The piece of
: DNA is prsent on the original pulse field gel at very low conc. but I use
: cybergreen instead of ethidium to stain as this is much more sensitive but
: i see nothing.  All help gratefully appreciated.

pH may be a problem w/ TBE & Qiaex.  Make sure that it is around 7.  I usually
add 1/10 volume of 1 M NaOAc, pH 6 to my extractions to lower the pH



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