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PLoS One. 2013 Oct 9;8(10):e76019. doi: 10.1371/journal.pone.0076019. eCollection 2013.

Efficient and rapid C. elegans transgenesis by bombardment and hygromycin B selection.

Author information

1
Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom.

Abstract

We report a simple, cost-effective, scalable and efficient method for creating transgenic Caenorhabditis elegans that requires minimal hands-on time. The method combines biolistic bombardment with selection for transgenics that bear a hygromycin B resistance gene on agar plates supplemented with hygromycin B, taking advantage of our observation that hygromycin B is sufficient to kill wild-type C. elegans at very low concentrations. Crucially, the method provides substantial improvements in the success of bombardments for isolating transmitting strains, the isolation of multiple independent strains, and the isolation of integrated strains: 100% of bombardments in a large data set yielded transgenics; 10 or more independent strains were isolated from 84% of bombardments, and up to 28 independent strains were isolated from a single bombardment; 82% of bombardments yielded stably transmitting integrated lines with most yielding multiple integrated lines. We anticipate that the selection will be widely adopted for C. elegans transgenesis via bombardment, and that hygromycin B resistance will be adopted as a marker in other approaches for manipulating, introducing or deleting DNA in C. elegans.

PMID:
24130756
PMCID:
PMC3794024
DOI:
10.1371/journal.pone.0076019
[Indexed for MEDLINE]
Free PMC Article

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