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Methods Digest, Vol 39, Issue 14 Large PCR p-GEM

Methods Digest, Vol 39, Issue 14 Large PCR p-GEM - Protocols and Methods Forum

Methods Digest, Vol 39, Issue 14 Large PCR p-GEM - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 08-23-2008, 02:56 AM
Jess
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Default Methods Digest, Vol 39, Issue 14 Large PCR p-GEM



The largest fragment I was ever able to clone was about 3 kB. I am not a cloning master, and maybe someone else managed to get a bigger one, at some point. We did not use Taq for the PCR, we used KOD polymerase to generate the PCR product, then A-tailed using Taq before ligation.

Cheers!

Jess * Grad. Res. Asst. University of Texas- Austin Institute for Cellular and Molecular Biology Department of Medicinal Chemistry ********** -Hook 'em Horns! *

--- On Thu, 8/21/08, [Only registered users see links. ] <[Only registered users see links. ].indiana.edu> wrote:
From: [Only registered users see links. ] <[Only registered users see links. ].indiana.edu>
Subject: Methods Digest, Vol 39, Issue 14
To: [Only registered users see links. ]
Date: Thursday, August 21, 2008, 12:06 PM

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Today's Topics:

1. Query about Maximum PCR product size for pGEM-T (Dario Dilernia)


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Message: 1
Date: Wed, 20 Aug 2008 13:01:10 -0300
From: "Dario Dilernia" <[Only registered users see links. ].ar>
Subject: Query about Maximum PCR product size for pGEM-T
To: <[Only registered users see links. ].indiana.edu>
Message-ID: <000b01c902dd$f687e960$e397bc20$@uba.ar>
Content-Type: text/plain; charset="iso-8859-1"

Dear Tony Schountz,



My name is Dario and I am from Argentina. I am sending you this mail because
I am looking for information regarding the maximum PCR fragment size that
can be cloned into pGEM-T Easy vector and I found on the internet that you
have asked the same question in a web forum although I didn´t find whether
somebody answered you… Did you find this information? It would be very
helpful for me. I see that your question was posted in the year 2000 so may
be you already have experience in this field. Did you have success cloning
large fragments with pGEM-T?



Best regards,



Dario









Lic. Dario A. Dilernia



Centro Nacional de Referencia para el SIDA

Departamento de Microbiología, Facultad de Medicina - UBA

Paraguay 2155 Piso 11º - C1121ABG

Buenos Aires, ARGENTINA

Tel: 54 11 4508-3671/3689 int: 132

Fax: 54 11 4508-3705






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