Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2000-06

AUTHORS

S Brenner, M Johnson, J Bridgham, G Golda, D H Lloyd, D Johnson, S Luo, S McCurdy, M Foy, M Ewan, R Roth, D George, S Eletr, G Albrecht, E Vermaas, S R Williams, K Moon, T Burcham, M Pallas, R B DuBridge, J Kirchner, K Fearon, J Mao, K Corcoran

ABSTRACT

We describe a novel sequencing approach that combines non-gel-based signature sequencing with in vitro cloning of millions of templates on separate 5 microm diameter microbeads. After constructing a microbead library of DNA templates by in vitro cloning, we assembled a planar array of a million template-containing microbeads in a flow cell at a density greater than 3x10(6) microbeads/cm2. Sequences of the free ends of the cloned templates on each microbead were then simultaneously analyzed using a fluorescence-based signature sequencing method that does not require DNA fragment separation. Signature sequences of 16-20 bases were obtained by repeated cycles of enzymatic cleavage with a type IIs restriction endonuclease, adaptor ligation, and sequence interrogation by encoded hybridization probes. The approach was validated by sequencing over 269,000 signatures from two cDNA libraries constructed from a fully sequenced strain of Saccharomyces cerevisiae, and by measuring gene expression levels in the human cell line THP-1. The approach provides an unprecedented depth of analysis permitting application of powerful statistical techniques for discovery of functional relationships among genes, whether known or unknown beforehand, or whether expressed at high or very low levels. More... »

PAGES

630-634

Journal

TITLE

Nature Biotechnology

ISSUE

6

VOLUME

18

Author Affiliations

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/76469

    DOI

    http://dx.doi.org/10.1038/76469

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/10835600


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    Download the RDF metadata as:  json-ld nt turtle xml License info

    HOW TO GET THIS DATA PROGRAMMATICALLY:

    JSON-LD is a popular format for linked data which is fully compatible with JSON.

    curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1038/76469'

    N-Triples is a line-based linked data format ideal for batch operations.

    curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1038/76469'

    Turtle is a human-readable linked data format.

    curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1038/76469'

    RDF/XML is a standard XML format for linked data.

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