Single Molecule Sequencing: Strategies and Technologies

This lecture covers the sequencing strategies and technologies used in single molecule sequencing, including Helicos' sequencing by synthesis method and Pacific Biosciences' real-time sequencing based on zero mode waveguides. It explains the process of adding fluorescent nucleotides one-by-one, cutting away the fluorescent molecule, and repeating the process until the DNA fragments are fully sequenced. The lecture also delves into the use of ZMWs, the incorporation of nucleotides by polymerase, and the diffusion of dye molecules. Additionally, it explores DNA translocation through nanopores, measuring current fluctuation during translocation, and the techniques of 'strand sequencing' and 'exonuclease sequencing' for real-time DNA sequencing.