Pacific Biosciences SMRT sequencing
Pacific Biosciences’ single molecule real time (SMRT) sequencing has some technical and conceptual advances over currently available massively parallel sequencing (MPS) technologies. It reduces sample manipulation and does not require amplification, thus potentially eliminating acquisition bias and keeping per sample costs low. Real-time measurement of base incorporation onto single template molecules theoretically allows for mutation detection at low levels, as well as detection of secondary structure and base modifications. Read lengths averaging 2,000 bp and growing far exceed those offered by other platforms. Errors, while frequent, occur in random locations and base composition, including homopolymers, pose no difficulties to obtaining sequence data. Low accuracy remains a limitation, but values are steadily improving and the use of circular consensus sequencing can overcome this limitation. PacBio currently shows promise as a supplemental sequencing system to assist in assembly of large genomes, as a rapid and affordable method to sequence small genomes and to explore microbiomes, and for mutation profiling and SNP discovery/confirmation in targeted genetic regions. LMT is working with the Sequencing Facility (SF) to test the strengths and weaknesses of the PacBio platform, target appropriate applications for PacBio sequencing, and ensure high quality performance prior to release as a CORE service. For more information about this novel sequencing platform visit the Pacific Biosciences website or click here for a detailed description of the technology.