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Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR).

ABSTRACT: A simple and efficient strategy for Bacterial Artificial Chromosome (BAC) recombineering based on co-selection is described. We show that it is possible to efficiently modify two positions of a BAC simultaneously by co-transformation of a single-stranded DNA oligo and a double-stranded selection cassette. The use of co-selection BAC recombineering reduces the DNA manipulation needed to make a conditional knockout gene targeting vector to only two steps: a single round of BAC modification followed by a retrieval step.

SUBMITTER: Newman RJ

PROVIDER: S-EPMC4627060 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature

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Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR).

Newman Robert J RJ Roose-Girma Merone M Warming Søren S

Nucleic acids research 20150618 19

A simple and efficient strategy for Bacterial Artificial Chromosome (BAC) recombineering based on co-selection is described. We show that it is possible to efficiently modify two positions of a BAC simultaneously by co-transformation of a single-stranded DNA oligo and a double-stranded selection cassette. The use of co-selection BAC recombineering reduces the DNA manipulation needed to make a conditional knockout gene targeting vector to only two steps: a single round of BAC modification followe ...[more]

PMID: 26089387

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