《Single-cell genomic sequencing using Multiple Displacement Amplification》.pdf

Single-cell genomic sequencing using Multiple Displacement Amplification Roger S Lasken Single microbial cells can now be sequenced using DNA without requiring development of culture methods. amplified by the Multiple Displacement Amplification (MDA) While amplification bias in the MDA reaction can result reaction. The few femtograms of DNA in a bacterium are in underrepresentation and loss of some sequences, the amplified into micrograms of high molecular weight DNA rewards are great often providing the first genomic suitable for DNA library construction and Sanger sequencing. sequences obtained from novel species. Furthermore, The MDA-generated DNA also performs well when used several studies have already achieved between 50 and directly as template for pyrosequencing by the 454 Life 75% completion of genomes from uncultured single cells, Sciences method. While MDA from single cells loses some of and method improvements are promising to improve the genomic sequence, this approach will greatly accelerate upon this further. Recent work has also demonstrated the pace of sequencing from uncultured microbes. The the power of single-cell sequencing to reveal the presence genetically linked sequences from single cells are also a of genes predicted from environmental and physiological powerful tool to be used in guiding genomic assembly of studies even from partial draft genomes. The genomic shotgun sequences of multiple organisms from environmental data could also assist in the process of eventually devel- DNA extracts (metagenomic sequences). oping culture methods, still a paramount goal required to Addresses