| description |
Assaying in vivo accrual of DNA damage and mutations by stem cells would further our understanding of aging and carcinogenesis. Two main hurdles hamper such assays. First, in vivo mutation rates are orders of magnitude lower than raw error rates of current sequencing procedures. Second, stem cells are vastly outnumbered by differentiated cells, which have a higher mutation rate — quantification of stem cell mutations is thus best performed from a small, well-defined stem cell population. Here we report a mutation detection technique, based on the “duplex sequencing” principle, with an estimated error rate below ~10-10 and that can start from as little as 50 pg DNA. We validate this technique using C. elegans germline stem cells, and characterize the dependence of damage and mutation accrual on cell cycling. Our technique provides the first direct measurements of in vivo, genome-wide mutation accrual of a stem cell population. |