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

[Jess]

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

Cheers!

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

--- On Thu, 8/21/08, methods-request from oat.bio.indiana.edu <methods-request from o=
at.bio.indiana.edu> wrote:
From: methods-request from oat.bio.indiana.edu <methods-request from oat.bio.indiana.=
edu>
Subject: Methods Digest, Vol 39, Issue 14
To: methods from magpie.bio.indiana.edu
Date: Thursday, August 21, 2008, 12:06 PM

Send Methods mailing list submissions to
=09methods from net.bio.net

To subscribe or unsubscribe via the World Wide Web, visit
=09http://www.bio.net/biomail/listinfo/methods
or, via email, send a message with subject or body 'help' to
=09methods-request from net.bio.net

You can reach the person managing the list at
=09methods-owner from net.bio.net

When replying, please edit your Subject line so it is more specific
than "Re: Contents of Methods digest..."


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 fmed.uba.ar>
Subject: Query about Maximum PCR product size for pGEM-T
To: <methods from magpie.bio.indiana.edu>
Message-ID: <000b01c902dd$f687e960$e397bc20$@uba.ar>
Content-Type: text/plain;=09charset=3D"iso-8859-1"

Dear Tony Schountz,

=20

My name is Dario and I am from Argentina. I am sending you this mail becaus=
e
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=
r
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?

=20

Best regards,

=20

Dario

=20

=20

=20

=20

Lic. Dario A. Dilernia

=20

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

=20

=20


--=20
Internal Virus Database is out-of-date.
Checked by AVG Free Edition.
Version: 7.5.432 / Virus Database: 268.16.2/613 - Release Date: 01/01/2007
02:50 p.m.
=20


------------------------------

_______________________________________________
Methods mailing list
Methods from net.bio.net
http://www.bio.net/biomail/listinfo/methods

End of Methods Digest, Vol 39, Issue 14
***************************************
=0A=0A=0A