Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Preparation of the A Homology Arm (A-Box) and B Homology Arm (B-Box)

(Protocol summary only for purposes of this preview site)

The 700-bp A homology arm (A-box) and the 700-bp B homology arm (B-box) are amplified by PCR using purified BAC DNA as template. For details on the primers used in these amplification reactions, see the information panel Primer Design for Homology Arms, Cointegration, and Resolution. The resulting A-box PCR product contains an AscI site at its 5 end (the 5 primer incorporates an AscI site, and the 3 primer does not incorporate any restriction sites). The B-box PCR product contains an XmaI site at its 3 end (the 5 primer does not incorporate any restriction sites, and the 3 primer incorporates an XmaI site). The amplification products are then digested with the appropriate restriction endonucleases to render them suitable for cloning into the shuttle vector (described in Protocol 6).


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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