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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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