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).
Lasken
Roger S
RS
J. Craig Venter Institute, 11149 N. Torrey Pines Rd., Suite 200, La Jolla, CA 92037, USA. rlasken@jcvi.org
D001419:Bacteria / Q000145:classification / Q000235:genetics*; D044822:Biodiversity; D004587:Electrophoresis, Agar Gel; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques