Cloning 5kb inserts

Don Walthers genejcky at yahoo.com
Wed Jan 24 14:43:11 EST 2001


Try cloning into a low copy vector into a MCS that is not downstream of a
promoter. With pACYC184 you can clone into one of the selectable markers and
pick some colonies to determine which are likely to have the insert.

--
Don Walthers  genejcky at yahoo.com
BS Microbiology U of Illinois at UC, 1996
PhD Microbiology U of Idaho, 2000
Our gene pool needs a little chlorine.
""Janakiraman Vasantharajan"" <vasant at PLANTPATH.KSU.EDU> wrote in message
news:3A655F8E.7598.3B007A at localhost...
> Hi netters,
> I had attempted to clone a 5kb insert (from a genomic library) into
> the bluescript vector atleast 3 times (into Kpn I - Sma I sites) with
> nosuccess. I am doing this only to sequence my inserts.  I do a blue
> white screening, none of my white colonies contain the insert. One
> of my colleagues suggested using the pUC 19 vector.
> Any suggestions ??
> You can email me at
> vasant at plantpath.ksu.edu
> Thanks in advance
>
>
> ---







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