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Two-Step Bacterial Artificial Chromosome (BAC) Engineering: Preparation and Verification of the Recombinant Shuttle Vector.

TitleTwo-Step Bacterial Artificial Chromosome (BAC) Engineering: Preparation and Verification of the Recombinant Shuttle Vector.
Publication TypeJournal Article
Year of Publication2020
AuthorsHeintz N, Gong S
JournalCold Spring Harb Protoc
Volume2020
Issue4
Pagination098061
Date Published2020 04 01
ISSN1559-6095
Abstract

Plasmid DNA is prepared from the recombinant shuttle vector pLD53.SCAB/A-B created by cloning of the A and B homology arms for two-step bacterial artificial chromosome (BAC) engineering. To confirm that the A-box and B-box arms have been successfully incorporated into pLD53.SCAB, the pattern of enzyme digestion of the modified plasmid is compared with that of the unmodified pLD53.SCAB. Once the shuttle vector is shown to carry the proper sequences, it is ready for transfer into the BAC host.

DOI10.1101/pdb.prot098061
Alternate JournalCold Spring Harb Protoc
PubMed ID32238592