Single-cell genomic sequencing using Multiple Displacement Amplification

Single-cell genomic sequencing using Multiple Displacement Amplification Single microbial cells can now be sequenced using DNA amplified by the Multiple Displacement Amplification (MDA) reaction. The few femtograms of DNA in a bacterium are amplified into micrograms of high molecular weight DNA suitable for DNA library construction and Sanger sequencing. The MDA-generated DNA also performs well when used directly as template for pyrosequencing by the 454 Life Sciences method. While MDA from single cells loses some of the genomic sequence, this approach will greatly accelerate the pace of sequencing from uncultured microbes. The genetically linked sequences from single cells are also a powerful tool to be used in guiding genomic assembly of shotgun sequences of multiple organisms from environmental DNA extracts (metagenomic sequences). http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Current Opinion in Microbiology Elsevier

Single-cell genomic sequencing using Multiple Displacement Amplification

Current Opinion in Microbiology, Volume 10 (5) – Oct 1, 2007

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Publisher
Elsevier
Copyright
Copyright © 2007 Elsevier Ltd
ISSN
1369-5274
DOI
10.1016/j.mib.2007.08.005
Publisher site
See Article on Publisher Site

Abstract

Single microbial cells can now be sequenced using DNA amplified by the Multiple Displacement Amplification (MDA) reaction. The few femtograms of DNA in a bacterium are amplified into micrograms of high molecular weight DNA suitable for DNA library construction and Sanger sequencing. The MDA-generated DNA also performs well when used directly as template for pyrosequencing by the 454 Life Sciences method. While MDA from single cells loses some of the genomic sequence, this approach will greatly accelerate the pace of sequencing from uncultured microbes. The genetically linked sequences from single cells are also a powerful tool to be used in guiding genomic assembly of shotgun sequences of multiple organisms from environmental DNA extracts (metagenomic sequences).

Journal

Current Opinion in MicrobiologyElsevier

Published: Oct 1, 2007

References

  • An improved method for single cell isolation of prokaryotes from meso-, thermo- and hyperthermophilic environments using micromanipulation
    Ishoy, T.; Kvist, T.; Westermann, P.; Ahring, B.K.
  • Specific single-cell isolation and genomic amplification of uncultured microorganisms
    Kvist, T.; Ahring, B.K.; Lasken, R.S.; Westermann, P.

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