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

Jess via (by shaakanin from
Fri Aug 22 21:56:00 EST 2008

The largest fragment I was ever able to clone was about 3 kB. I am not a cl=
oning master, and maybe someone else managed to get a bigger one, at some p=
oint. We did not use Taq for the PCR, we used KOD polymerase to generate th=
e PCR product, then A-tailed using Taq before ligation.=20


Jess  =A0  Grad. Res. Asst.  University of Texas- Austin  Institute for Cel=
lular and Molecular Biology  Department of Medicinal Chemistry  =A0=A0=A0=
=A0=A0=A0=A0=A0=A0=A0 -Hook 'em Horns!  =A0

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From: methods-request from <methods-request from
Subject: Methods Digest, Vol 39, Issue 14
To: methods from
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)


Message: 1
Date: Wed, 20 Aug 2008 13:01:10 -0300
From: "Dario Dilernia" <ddilernia from>
Subject: Query about Maximum PCR product size for pGEM-T
To: <methods from>
Message-ID: <000b01c902dd$f687e960$e397bc20$>
Content-Type: text/plain;=09charset=3D"iso-8859-1"

Dear Tony Schountz,


My name is Dario and I am from Argentina. I am sending you this mail becaus=
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=B4t find whethe=
somebody answered you=85 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,







Lic. Dario A. Dilernia


Centro Nacional de Referencia para el SIDA

Departamento de Microbiolog=EDa, Facultad de Medicina - UBA

Paraguay 2155 Piso 11=BA - C1121ABG=20

Buenos Aires, ARGENTINA

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

Fax: 54 11 4508-3705



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