Characteristics of 454 pyrosequencing data—enabling realistic simulation with flowsim
Bioinformatics, 26 (18):
i420-i425 (2010)DOI: 10.1093/bioinformatics/btq365
Abstract
Motivation: The commercial launch of 454 pyrosequencing in 2005 was a milestone in genome sequencing in terms of performance and cost. Throughout the three available releases, average read lengths have increased to ∼500 base pairs and are thus approaching read lengths obtained from traditional Sanger sequencing. Study design of sequencing projects would benefit from being able to simulate experiments.Results: We explore 454 raw data to investigate its characteristics and derive empirical distributions for the flow values generated by pyrosequencing. Based on our findings, we implement Flowsim, a simulator that generates realistic pyrosequencing data files of arbitrary size from a given set of input DNA sequences. We finally use our simulator to examine the impact of sequence lengths on the results of concrete whole-genome assemblies, and we suggest its use in planning of sequencing projects, benchmarking of assembly methods and other fields.Availability: Flowsim is freely available under the General Public License from http://blog.malde.org/index.php/flowsim/Contact: susanne.balzer@imr.no; ketil.malde@imr.no
