Data from: Benefits and challenges with applying unique molecular identifiers in next generation sequencing to detect low frequency mutations
Cite this dataset
Kou, Ruqin et al. (2016). Data from: Benefits and challenges with applying unique molecular identifiers in next generation sequencing to detect low frequency mutations [Dataset]. Dryad. https://doi.org/10.5061/dryad.n6068
Indexing individual template molecules with a unique identifier (UID) before PCR and deep sequencing is promising for detecting low frequency mutations, as true mutations could be distinguished from PCR errors or sequencing errors based on consensus among reads sharing same index. In an effort to develop a robust assay to detect from urine low-abundant bladder cancer cells carrying well-documented mutations, we have tested the idea first on a set of mock templates, with wild type and known mutants mixed at defined ratios. We have measured the combined error rate for PCR and Illumina sequencing at each nucleotide position of three exons, and demonstrated the power of a UID in distinguishing and correcting errors. In addition, we have demonstrated that PCR sampling bias, rather than PCR errors, challenges the UID-deep sequencing method in faithfully detecting low frequency mutation.