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  Novel method for high-throughput colony PCR screening in nanoliter-reactors.

Walser, M., Pellaux, R., Meyer, A., Bechtold, M., Vanderschuren, H., Reinhardt, R., et al. (2009). Novel method for high-throughput colony PCR screening in nanoliter-reactors. Nucleic Acids Research, 37(8), e57-e57. doi:10.1093/nar/gkp160.

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Genre: Journal Article
Alternative Title : Nucleic Acids Res.

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 Creators:
Walser, Marcel, Author
Pellaux, Rene, Author
Meyer, Andreas, Author
Bechtold, Matthias, Author
Vanderschuren, Herve, Author
Reinhardt, Richard1, Author           
Magyar, Joseph, Author
Panke, Sven, Author
Held, Martin, Author
Affiliations:
1High Throughput Technologies, Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433552              

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 Abstract: We introduce a technology for the rapid identification and sequencing of conserved DNA elements employing a novel suspension array based on nanoliter (nl)-reactors made from alginate. The reactors have a volume of 35 nl and serve as reaction compartments during monoseptic growth of microbial library clones, colony lysis, thermocycling and screening for sequence motifs via semi-quantitative fluorescence analyses. nl-Reactors were kept in suspension during all high-throughput steps which allowed performing the protocol in a highly space-effective fashion and at negligible expenses of consumables and reagents. As a first application, 11 high-quality microsatellites for polymorphism studies in cassava were isolated and sequenced out of a library of 20 000 clones in 2 days. The technology is widely scalable and we envision that throughputs for nl-reactor based screenings can be increased up to 100 000 and more samples per day thereby efficiently complementing protocols based on established deep-sequencing technologies.

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Language(s): eng - English
 Dates: 2009-03-12
 Publication Status: Issued
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Title: Nucleic Acids Research
  Alternative Title : Nucleic Acids Res.
Source Genre: Journal
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Pages: - Volume / Issue: 37 (8) Sequence Number: - Start / End Page: e57 - e57 Identifier: ISSN: 0305-1048