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

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Using Multisite LR Cloning to Generate a Destination Clone

(Protocol summary only for purposes of this preview site)

A multisite cloning reaction transfers the inserts from multiple Entry clones into a single Destination vector (Fig. 4 in the chapter introduction). This type of recombination is much less efficient than transferring a single DNA fragment; however, the variety of Destination clones that can be generated in this manner is vast. In this example protocol, we describe using pDEST-MB14 (Dupuy et al. 2007) to make a Destination clone that features a promoter fragment fused upstream to an ORF that is cloned in-frame with a carboxy-terminal GFP moiety encoded by the plasmid backbone. Just as with Protocol 2, this method can be used as a guide for other multisite cloning reactions.


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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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