Abstract

BackgroundSeveral standalone error correction tools have been proposed to correct sequencing errors in Illumina data in order to facilitate de novo genome assembly. However, in a recent survey, we showed that state-of-the-art assemblers often did not benefit from this pre-correction step. We found that many error correction tools introduce new errors in reads that overlap highly repetitive DNA regions such as low-complexity patterns or short homopolymers, ultimately leading to a more fragmented assembly.ResultsWe propose BrownieCorrector, an error correction tool for Illumina sequencing data that focuses on the correction of only those reads that overlap short DNA patterns that are highly repetitive in the genome. BrownieCorrector extracts all reads that contain such a pattern and clusters them into different groups using a community detection algorithm that takes into account both the sequence similarity between overlapping reads and their respective paired-end reads. Each cluster holds reads that originate from the same genomic region and hence each cluster can be corrected individually, thus providing a consistent correction for all reads within that cluster.ConclusionsBrownieCorrector is benchmarked using six real Illumina datasets for different eukaryotic genomes. The prior use of BrownieCorrector improves assembly results over the use of uncorrected reads in all cases. In comparison with other error correction tools, BrownieCorrector leads to the best assembly results in most cases even though less than 2% of the reads within a dataset are corrected. Additionally, we investigate the impact of error correction on hybrid assembly where the corrected Illumina reads are supplemented with PacBio data. Our results confirm that BrownieCorrector improves the quality of hybrid genome assembly as well. BrownieCorrector is written in standard C++11 and released under GPL license. BrownieCorrector relies on multithreading to take advantage of multi-core/multi-CPU systems. The source code is available at https://github.com/biointec/browniecorrector.

Highlights

  • Illumina platforms generate accurate sequencing data with high throughput at a low financial cost

  • BrownieCorrector performs worse than Karect in datasets D4 and D7 which is due to the fact that the standard deviation for the R4 Illumina dataset is 92, which is relatively high compared to the other datasets

  • BrownieCorrector uses the entire read sequence as well as the paired-end read information to cluster read pairs in homogeneous groups, where the paired-end reads in each group originate from the same genomic region

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Summary

Introduction

Illumina platforms generate accurate sequencing data with high throughput at a low financial cost. It is estimated that more than 90% of sequencing data worldwide are generated by Illumina platforms These data are characterized by a relatively short read length (100-300 bp) and low error rate (1-2% errors). As a single sequencing error leads to up to k erroneous k-mers in the DBG, true nodes in the DBG are vastly outnumbered by erroneous nodes. These artifacts highly complicate the task of identifying the path in the graph that represents the original genomic sequence. We found that many error correction tools introduce new errors in reads that overlap highly repetitive DNA regions such as low-complexity patterns or short homopolymers, leading to a more fragmented assembly

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