Re: [ccp4bb] Expression vector with NdeI-ClaI sites

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Expression vector with NdeI-ClaI sites
From: "Dunten, Pete W {- dot -} " pete {- at -} SLAC {- dot -} STANFORD {- dot -} EDU
Date: 2008-07-21

Next message:
Subject: Spooky, moving crystals
From: "Mark J {- dot -} van Raaij" mark {- dot -} vanraaij {- at -} USC {- dot -} ES
Date: 2008-07-21

Subject: Re: Expression vector with NdeI-ClaI sites
From: Mark Collins mnc2003 {- at -} COLUMBIA {- dot -} EDU
Date: 2008-07-21

Or do the new school cloning, SLIC (Sequence & Ligation Independent
Cloning) and "subclone" into any position in a vector.
This method uses a PCR product and vector, the PCR primers have 20-40bp
overlap with a region in the vector. Mix cut and purified vector with PCR
product. Digest with T4 polymerase, quench, and transform.
When I do the PCR in the morning I have clonies the next day.

REF:
Harnessing homologous recombination in vitro to generate recombinant DNA
via SLIC
Mamie Z Li & Stephen J Elledge
Nat Meth V4(3) pp 251

Mark

------------------------------------------------------------------------------
Mark Collins
Columbia University
Biochemistry & Molecular Biophysics
Black Building 259/201 Office/Lab
212 305 1951 (work)
mnc2003@columbia.edu

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Expression vector with NdeI-ClaI sites
From: "Dunten, Pete W {- dot -} " pete {- at -} SLAC {- dot -} STANFORD {- dot -} EDU
Date: 2008-07-21

Next message:
Subject: Spooky, moving crystals
From: "Mark J {- dot -} van Raaij" mark {- dot -} vanraaij {- at -} USC {- dot -} ES
Date: 2008-07-21