Dr Francisco M. De La Vega

Title: The road to 99.999% accuracy single molecule sequencing

Life Technologies continues to improve the SOLiD™ System, a massively parallel second-generation sequencing system based on oligonucleotide probe ligation and clonally amplified DNA fragments attached to beads. The use of beads have provided increasing sequencing throughputs reaching 100 Gb/run today, and soon approaching 300 Gb/run by increasing packing and reducing bead dimension coupled to signal to noise improvements. Recent chemistry enhancements allow longer read lengths, providing 25-125 bp reads from either paired-end or mate-pair libraries with insert sizes ranging from ~100bp to ~20 kb. This ever-increasing throughput has reduced the cost of sequencing a human genome to US$6,000 today, to US$3,000 by the end of 2010, and continues to come down. A cornerstone of the SOLiD technology is its error detection and correction ability. Currently in SOLiD 4, error detection and reference-assisted correction codes provides an average 99.95% read accuracy, the lowest in second-generation sequencing. We are working in new error corrections codes for the SOLiD System that can provide up to 99.999% read accuracy without the need of reference sequence, which would be essential in applications such as somatic mutation detection, sequencing pooled or complex samples, and low pass GWAS by sequencing, among others. In addition, Life Technologies is actively developing a novel third generation single-molecule sequencing platform based on monitoring FRET on the surface of a quantum-dot (Qdot®nanocrystals). This technology provides a portable nanometer-sized sequencing engine that enables continuous long read lengths using active-sequencer exchange and tunable high accuracy using recursive sequencing. The ability to rapidly and inexpensively produce very long reads can enable new applications that rely on understanding the long-range structure of nucleic acids and, in combination with highly accurate “short-read” SOLiD System data, would provide flexible and comprehensive genome analysis solutions.