Re: [DIYbio] Re: SLICE -- Seamless Ligation Cloning Extract... or, using lambda phage homologous recombination in-vitro

Our lab uses the PPY strain (a generous gift from Einstein U) for our cloning needs. We exactly follow their prescribed protocols to prep the extract, buffer, etc.

This method is a true gem. Side by side with Gibson, it works at least as well and often better. For basic insertion of genes into vectors we obtain lawns of cells (too many colonies sometimes), and all tested colonies are colony PCR positive and sequencing comes back as intended.

Economically it is very difficult to argue using Gibson over PPY-based SLiCE cloning.




On Thursday, October 17, 2013 at 3:05:20 PM UTC-7, Andrew wrote:
I too got this strain and no luck, but I'm no pro.  Anyone else have trouble or success?  A recent article on synthetic assembly methods stated that of survey respondents, around 10%, or 13 respondents, used SLICE.  I might try CPEC instead, and I really want ABI-REC to work, but so far no luck...

On Wednesday, September 18, 2013 4:08:05 PM UTC-7, Nathan McCorkle wrote:
On Thu, Sep 12, 2013 at 2:54 PM, yamwa <yam...@gmail.com> wrote:
> Did anyone try out this method? I got the PPY strain from Einstein but don't
> have luck with it :/

Didn't have luck in what sense? How long were your homologous flanking regions?

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