Transgenic mice vector construct

Pow Joshi via methods%40net.bio.net (by pow.joshi from gmail.com)
Tue Dec 4 14:51:58 EST 2007


On 04/12/2007, rosaak <rosaak from gmail.com> wrote:
>
> Hi all,
>
> I have constructed a vector consting cDNA for making transgenic mice.
> The backbone is pCI contining CMV promoter.For microinjections i need
> to linearise the vector., and there are no suitable sites for it.
>
> My doubt is that can i use a set of primers to pcr amplify CMVpromoter-
> cDNA-polyA region from the construct and use it for microinjection.
> [i'll be using a highfidility enzyme for pcr.]
>
> Is it techically ok to do this.If so how will i purfy it ?



well,  I cannot say anything about the pcr,

for purification, I use the Qiagen kit, and it works quite well, at least
for my purposes.... but then Iuse zebrafish eggs.
  You could also use the standard phenol chloroform extraction foillowed ny
isopropanol/ ethanol precipitation.

Hope that helps

Pow



Please reply
>
> regards,
>
> RP
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