Cotransformation
Aawara Chowdhury
via methods%40net.bio.net
(by aawara from FEMA-trailer.org)
Tue Jun 5 20:19:38 EST 2007
In <mailman.1361.1181055370.5139.methods from net.bio.net>,
Pow Joshi <pow.joshi from gmail.com> wrote:
> Transfecting both plasmids together has worked rather well for me in the
> past, especially with transient transfections ..... you would need to
> optimize the concentrations of each .....
The original poster wasn't attempting a transient transfection of animal
cells. He wanted to co-transform two plasmids into BL21(DE3), a strain
of E. coli.
AC
>
> On 6/4/07, waikhay <waikhay from gmail.com> wrote:
>>
>> Hi all! :)
>>
>> I would like to transform 2 plasmids i.e. an expression vector (carrying
>> the
>> gene of interest) and a reporter vector (into which the promoter of
>> interest
>> has been cloned) into BL21(DE3) cells. Would you recommend/suggest
>> transforming both plasmids into the cells at once (cotransform) or
>> transforming one plasmid after another (i.e. transform the reporter vector
>> into the cells first, select for transformants, then only transform the
>> expression vector and select for cotransformants)?
>>
>> Thanks much,
>> Waikhay
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