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

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Two-Step BAC Engineering: Preparation of Shuttle Vector DNA

(Protocol summary only for purposes of this preview site)

In the two-step approach, a single plasmid is introduced into the BAC-carrying cell lines (see Figs. 4 and 5 in the chapter introduction). The shuttle vector pLD53.SCAB (or pLD53.SCAEB) (see Fig. 5 in the chapter introduction) carries the recA gene as well as the R6K origin, which requires the protein to replicate. PIR2 cells, expressing , are typically used for the amplification of the vector and maintain about 15 copies/cell of the donor vector, which is relatively stable in this host.


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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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