How to use Expand PCR System

Agustin Alconada alconada at embl-heidelberg.de
Wed Sep 18 08:32:50 EST 1996


In article <199609161450.WAA08685 at public.cc.jl.cn>, ouyhs at PUBLIC.CC.JL.CN wrote:

> Dear Netter:
> 
>     I have bought a Expand Long Template PCR System and a Expand High
> Fidelity PCR System From Boehringer Mannheim. I wish I could use
> these kits to amplify and clone a gene about 10kb from bovine genome 
> DNA. But I do not know which kind end be produced in the PCR products
> if we use those Expand systems.  Can I use the T-vctor to clone the
> PCR products? Do you have any literature on amplifing and cloning 
> the DNA by using those Expand systems?
> 

   I use regularly the Expand High Fidelity PCR system, but I have never
tried to amplify anything longer than 1.5 kb. Although the Boehringer
salesman claimed that it is not possible to clone PCR products generated
with the Expand system into T-vectors, I have been doing it for a few
months without any problem. I use the TA cloning kit from Invitrogen and
always find between 2 and 10 white colonies, which are all positive.

_____________________________________________________________________Agustin Alconada
European Molecular Biology Laboratory
Meyerhofstrasse 1
69117 Heidelberg
Phone:  49-6221-387286
Fax:    49-6221-387306

http://www.embl-heidelberg.de/~alconada
_____________________________________________________________________



More information about the Methods mailing list