Abstract

BackgroundThe third generation PacBio SMRT long reads can effectively address the read length issue of the second generation sequencing technology, but contain approximately 15% sequencing errors. Several error correction algorithms have been designed to efficiently reduce the error rate to 1%, but they discard large amounts of uncorrected bases and thus lead to low throughput. This loss of bases could limit the completeness of downstream assemblies and the accuracy of analysis.ResultsHere, we introduce HALC, a high throughput algorithm for long read error correction. HALC aligns the long reads to short read contigs from the same species with a relatively low identity requirement so that a long read region can be aligned to at least one contig region, including its true genome region’s repeats in the contigs sufficiently similar to it (similar repeat based alignment approach). It then constructs a contig graph and, for each long read, references the other long reads’ alignments to find the most accurate alignment and correct it with the aligned contig regions (long read support based validation approach). Even though some long read regions without the true genome regions in the contigs are corrected with their repeats, this approach makes it possible to further refine these long read regions with the initial insufficient short reads and correct the uncorrected regions in between. In our performance tests on E. coli, A. thaliana and Maylandia zebra data sets, HALC was able to obtain 6.7-41.1% higher throughput than the existing algorithms while maintaining comparable accuracy. The HALC corrected long reads can thus result in 11.4-60.7% longer assembled contigs than the existing algorithms.ConclusionsThe HALC software can be downloaded for free from this site: https://github.com/lanl001/halc.

Highlights

  • The third generation PacBio SMRT long reads can effectively address the read length issue of the second generation sequencing technology, but contain approximately 15% sequencing errors

  • Experimental design To evaluate the performance of HALC, we ran HALC on three data sets from the species, E. coli, A. thaliana and Maylandia zebra, of small, medium and large genome sizes, respectively

  • Except for PacBioToCA and LSC, the average read length of all the algorithms is inversely proportional to the throughput because more but shorter reads can be obtained with higher throughput

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Summary

Introduction

Several error correction algorithms have been designed to efficiently reduce the error rate to 1%, but they discard large amounts of uncorrected bases and lead to low throughput This loss of bases could limit the completeness of downstream assemblies and the accuracy of analysis. A tremendous number of species have been assembled from short reads, but most of the assemblies are incomplete and fragmented into several thousands of contigs [2, 3] To address this issue, the PacBio SMRT sequencing technology, as a representative of third generation sequencing technology, has been attracting more and more attention since its commercial release in 2010 [4]. Depending on how the long reads are used, sequencing projects can be grouped into two classes

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