KI and DNA purification

jag100 at snethen.com jag100 at snethen.com
Mon Mar 29 19:27:50 EST 1999


I have inherited a protocol for purifying DNA from agarose gels with
Promega's AgarACE enzyme.  The protocol I have calls for melting the gel
slices in 5.2M KI (final conc ~1.7M in ~300 ul total volume) before adding
the AgarACE.

I know KI lowers melting and gelling temps, but I can't find any reference as
to how much I should use.  The Promega protocol doesn't call for it, so I am
a bit at a loss to explain its presence, or why I need what seems like a high
concentration.	Also, I started storing it in plastic flasks; because it had
been stored in glass bottles, someone in my group wondered if there was a
good reason for this.

I am still working on getting away from AgarACE altogether, so this might be
a moot point.  But, for curiosity's sake, I would appreciate any comments the
group has on KI.  I apologise if this is a basic biochem/molbio question--I
looked in Maniatis and my biochem book, plus a quick search on the web, and
found nothing of use.

Thanks,

Jennifer
jag100 at snethen.com

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