Small-scale extraction of Caenorhabditis elegans genomic DNA

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dc.contributor.authorMadhu, Bhoomi
dc.contributor.authorLakdawala, Mohammed Farhan
dc.contributor.authorGumienny, Tina L.
dc.creator.orcidhttps://orcid.org/0000-0002-3932-7815
dc.date.accessioned2023-07-26T18:56:16Z
dc.date.available2023-07-26T18:56:16Z
dc.date.issued2022
dc.description.abstractGenomic DNA extraction from single or a few Caenorhabditis elegans has many downstream applications, including PCR for genotyping lines, cloning, and sequencing. The traditional proteinase K-based methods for genomic DNA extraction from C. elegans take several hours. Commercial extraction kits that effectively break open the C. elegans cuticle and extract genomic DNA are limited. An easy, faster (~15 min), and cost-efficient method of extracting C. elegans genomic DNA that works well for classroom and research applications is reported here. This DNA extraction method is optimized to use single or a few late-larval (L4) or adult nematodes as starting material for obtaining a reliable template to perform PCR. The results indicate that the DNA quality is suitable for amplifying gene targets of different sizes by PCR, permitting genotyping of single or a few animals even at dilutions to one-fiftieth of the genomic DNA from a single adult per reaction. The reported protocols can be reliably used to quickly produce DNA template from a single or a small sample of C. elegans for PCRbased applications.en_US
dc.description.sponsorshipThe N2 strain and E. coli OP50 bacteria were obtained from the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). The blmp-1(tm548) strain was obtained from the National Bioresource Project, Japan. The authors thank WormBase. This work was supported by NIH R01GM097591 to T.L.G., internal funding by Texas Woman's University to T.L.G, and TWU's Center for Student Research to M.F.L.en_US
dc.identifier.citationThis is a published version of an article that is available at: https://doi.org/10.3791/63716-v. Recommended citation: Madhu, B., Lakdawala, M. F., & Gumienny, T. (2022). Small-scale extraction of Caenorhabditis elegans genomic DNA. Journal of Visualized Experiments, (184), e63716. This item has been deposited in accordance with publisher copyright and licensing terms and with the author’s permission.en_US
dc.identifier.urihttps://hdl.handle.net/11274/15272
dc.identifier.urihttps://doi.org/10.3791/63716-v
dc.language.isoen_USen_US
dc.publisherJOVEen_US
dc.rights.holder© 2022 JoVE Journal of Visualized Experiments
dc.subjectGenomic DNA extractionen_US
dc.subjectCost-efficienten_US
dc.subjectPCRen_US
dc.titleSmall-scale extraction of Caenorhabditis elegans genomic DNAen_US
dc.typeArticleen_US

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