Friday, May 15, 2009, 4:00 pm — John Dunn Seminar Room, Bldg. 463
The majority (>90%) of a prokaryotic genome can be assembled de novo from 454 FLX shotgun reads derived from an individual cell. We are working to establish a general-purpose pipeline for single-cell genomics applicable to any cell that can be visualized under a microscope, including rare members of complex microbial consortia. We have developed an integrated microfluidic device that can carry out 32 independent, low-bias multiple displacement amplification (MDA) reactions on genomic DNA templates derived from single cells. High-performance optical trapping of cells inside the device enables rapid (up to 1 cell per minute), definitive sorting of single cells from high-concentration cell suspensions, and stringent exclusion of contaminates from the nanoliter-scale amplification reactions. Sequencing of sub-nanogram DNA samples without pre-amplification will be discussed, along with strategies for genome assembly addressing representation bias and chimerism in the raw sequencing reads.
Hosted by: Wally Mangel
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