Nested Deletions Kits for Sequencing

Kimberley Snowden kcsnowden at ucdavis.edu
Mon Oct 7 13:08:47 EST 1996


Another alternative is the DNAse I deletion strategy, which has worked
well for me.  It doesn't require a kit at all.  A reference for this
method is as follows:
Lin et al., 1985  Analytical Biochemistry 147: 114-119
The only thing I changed about this method was that I substituted all
the precipitation steps with clean ups using wizard/magic columns (no
affiliation with Promega, and many other companies products would work
equally as well) which increased my yield about 1000-fold.
Hope this helps.
Kim Snowden
kcsnowden at ucdavis.edu


martinez at puccini.CRL.umn.edu (J. Pat Martinez) wrote:

>I need to sequence a 10 kb fragment and will use a strategy based on a
>series of nested deletions.  I'm aware of two different strategies for
>generating nested deletions.  One strategy  uses Exonuclease III and
>Mungbean nuclease, and the other uses transposon mediated deletions
>(Deletion factory by BRL).  I was leaning toward the Exonuclease/Mungbean
>nuclease based kits.   Any suggestions on which manufacture to go with,
>such as Promega's Erase a Base kit or Stratagene's deletion kit.  Thanks
>for your suggestions.

>Pat M.





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