Risks of double-counting in deep sequencing

Lou et al. (1) report a technique for increased sensitivity of DNA sequencing that they call “circle sequencing.” The authors’ method involves circularizing single-stranded DNA and performing rolling-circle amplification (RCA). Multiple duplicates are thereby generated, and errors can be removed by comparing the sequences of the duplicates. Lou et al. report accuracy similar to that of single-stranded tagging techniques (2, 3), but less than that achieved with double-stranded tagging (4). The authors propose their method as a high-efficiency tool for deep-sequencing of heterogeneous DNA samples.

[1]  Jeffrey A. Hussmann,et al.  High-throughput DNA sequencing errors are reduced by orders of magnitude using circle sequencing , 2013, Proceedings of the National Academy of Sciences.

[2]  Jesse J. Salk,et al.  Detection of ultra-rare mutations by next-generation sequencing , 2012, Proceedings of the National Academy of Sciences.

[3]  Cassandra B. Jabara,et al.  Accurate sampling and deep sequencing of the HIV-1 protease gene using a Primer ID , 2011, Proceedings of the National Academy of Sciences.

[4]  K. Kinzler,et al.  Detection and quantification of rare mutations with massively parallel sequencing , 2011, Proceedings of the National Academy of Sciences.