TA cloning of a PCR product generate by Vent DNA polymerase

Bo Pontoppidan bo.pontoppidan at cellfo.slu.se
Wed Jul 30 11:54:22 EST 1997


Dr. Yu wrote:
> 
> Does anyone has any successful experience in cloning a PCR product
> generated by Vent DNA polymerase? The product is 2.7 kbp.  I followed
> the
> InVitrogen protocol and incubate the Vent PCR product in Taq at 72 degC
> for 10 minutes, phenol extract and preciptate the DNA at room temp. The
> precipitated DNA is resuspended in TE and immediate ligate to the TA
> vector. I tried this several times but I don't get any clones.
> 
Try to do a blunt end ligation directly with the vent pcr product.

Bo

--
Bo Pontoppidan                                  Phone: (+46) 018 67 33
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Uppsala Genetic Center                            Fax: (+46) 018 67 32
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Dept. of Cell Research                     Home Phone: (+46) 018 69 28
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Swedish University of Agricultural Sciences, Box 7055,  
S-750 07 Uppsala, Sweden             E-mail:
bo.pontoppidan at cellfo.slu.se



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