de.mpg.escidoc.pubman.appbase.FacesBean
English
 
Help Guide Disclaimer Contact us Login
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

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

MPS-Authors
http://pubman.mpdl.mpg.de/cone/persons/resource/persons50488

Reinhardt,  Richard
High Throughput Technologies, Max Planck Institute for Molecular Genetics, Max Planck Society;

Locator
There are no locators available
Fulltext (public)

e57.pdf
(Any fulltext), 5MB

Supplementary Material (public)
There is no public supplementary material available
Citation

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.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0010-7DED-5
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.