Mutation assay using SMRT DNA sequencer

Assessment of very low frequency of somatic mutations is important in the research fields of environmental mutagenesis, carciongenesis, epidemiology, and regulatory science. Recently, the PacBio RS II DNA sequencer which is based on a single molecule real time (SMRT) technology became available. Since it can read DNA sequence in each single DNA molecules, in principle, it is able to detect mutations present in the molecules by just sequencing it accurately. To ensure accuracy of the sequence data, "SMRTbell template" in which single stranded DNA loops are ligated to both ends of a double stranded DNA , is used and sequence the same DNA molecules repeatedly. In this study, we validated if we can detect very low frequency of mutation by this strategy.