August 19, 2013  |  Products, procedures + protocols

Scientists Assess Error Modes in Sequencing Platforms and Find SMRT Sequencing ‘Least Biased’

A paper from scientists at the Broad Institute reports a rigorous study of bias across all major sequencing platforms. In “Characterizing and measuring bias in sequence data,” published in Genome Biology, lead author Michael Ross and his colleagues report that SMRT® Sequencing on the PacBio® sequencer is the “least biased” in coverage of all the technologies studied.

The authors assessed sequences for coverage bias, or uniformity of read distribution, and error bias, or incorrect call at a given position. For coverage bias, they report that PacBio performed best in extreme GC content (both GC-rich and GC-poor) and suggest this may be related to the lack of an amplification step in the sequencing process. Regarding error bias, the scientists describe shifting error rates based on genome sequence; GC-rich or homopolymer regions, for example, tended to change the rate of errors for each platform. “In general, the sequence context dependence of error rates varied considerably from technology to technology,” they write.

Ross et al. note that each platform’s bias rate changes with technology development, but note that at the time of their work, “single-molecule data from Pacific Biosciences” had “the clear edge.”

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