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.
kcsnowden at ucdavis.edumartinez 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.