T/A cloning again. To Kinase or Not
rafael at howard.genetics.utah.edu
Wed Jan 17 19:41:09 EST 1996
On 17 Jan 1996, HIRANYA ROYCHOWDHURY wrote:
> On 16 Jan 1996, Francois-Xavier Maquart wrote:
> > Dear Netters.
> > I have just discover this newsgroup. Fascinating.
> > At the begining I have a naive question: Should I kinase PCR products
> > (or oligos)
> Yes, phosphorylating will definitely better the chance.
> for T/A cloning? Either Promega pGEM-T vector system kit
> > or Invitrogen TA cloning system kit do not discuss this problem.
I don't get the point. Kinase what? PCR product, as it is, is already
kinased (I mean that it has a phosphate at the 5' end).
Oligos are different. You have to make sure your oligos are kinased for
cloning, but it is not strictily necesary if your vector is kinased (or,
it is not dephosphatased). But only in the case of DIRECT cloning of
oligos, not through PCR.
Rafael Maldonado | La cita ha sido
room 6160 Eccles Institute of Human Genetics |
Department of Human Genetics | retirada por respeto
University of Utah |
Salt Lake City, Utah 84112. USA. | a la propiedad
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