E.coli strain recommendation, please
Brian P Higgins
(by higginsb78 from gmail.com)
Wed Jun 18 11:45:41 EST 2008
I'll second DH5alpha--works very well. I'd also suggest looking at your NEB
catalog--they have several E. coli strains available for free (you pay
shipping if you're not ordering something else). I know they offer several
of the JM10X's, which are very handy (especially the recA strain, which is
what you'd want). There are too many alternatives to list, so you'll have
to be a bit more specific about the desired genotype for me to give more
I am not sure why you're having the problem with Top10. I've used it many
times with great success and I have not found it to grow slowly at all.
Could it be the level of antibiotic you are using?
On Wed, Jun 18, 2008 at 11:51 AM, DK <dk from no.email.thankstospam.net> wrote:
> Traditionally, for reasons I am not quite sure about, we use
> Invitrogen's TOP10 cells for all routine cloning. There is one
> serious problem with this strain, however. Cells grow too slowly.
> E.g. to get a decent size clone, plates need to be incubated
> 24 hours. In a typical overnight (4 pm - 9 am), clones are too
> small. Also, "short" overnight liquid cultures are not saturated,
> resulting in low plasmid yields. This is becoming a problem
> when we want to work fast and not waste time waiting for the
> bugs to grow. .
> So I am looking for alternatives that offer:
> 1. High transformation efficiency (both chemical
> and electroporation).
> 2. Low level of recombination.
> 3. High plasmid yields and quality of DNA minipreps.
> 4. Fast growth.
> TOP10 fit requirements #1-2, #3 if we wait long enough
> but definitely not #4.
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