Improving the performance of true-Single Molecule Sequencing for ancient DNA

Helicos true Single-Molecule DNA Sequencing (tSMS) has recently shown great potential for sequencing DNA molecules from Pleistocene fossils. Here we test the performance of two novel tSMS template preparation methods on ancient DNA. We demonstrate that DNA phosphatase treatments and mild denaturation temperatures increase the amount of sequence information recovered from ancient bones. Together with molecular preservation niches identified in large bone crystals, these procedures could be used to increase sequence coverage of ancient genomes.