Hybrid assembly of the large and highly repetitive genome of Aegilops tauschii, a progenitor of bread wheat, with the mega-reads algorithm
- Aleksey V. Zimin1,
- Daniela Puiu2,
- Ming-Cheng Luo3,
- Tingting Zhu3,
- Sergey Koren4,
- Guillaume Marcais5,
- James A. Yorke1,
- Jan Dvorak3 and
- Steven L. Salzberg2,6
- 1 University of Maryland;
- 2 Johns Hopkins University;
- 3 University of California, Davis;
- 4 National Human Genome Research Institute, NIH;
- 5 Carnegie Mellon University
- ↵* Corresponding author; email: salzberg{at}jhu.edu
Abstract
LLong sequencing reads generated by single-molecule sequencing technology offer the possibility of dramatically improving the contiguity of genome assemblies. The biggest challenge today is that long reads have relatively high error rates, currently around 15%. The high error rates make it difficult to use this data alone, particularly with highly repetitive plant genomes. Errors in the raw data can lead to insertion or deletion errors (indels) in the consensus genome sequence, which in turn create significant problems for downstream analysis; for example, a single indel may shift the reading frame and incorrectly truncate a protein sequence. Here we describe an algorithm that solves the high error rate problem by combining long, high-error reads with shorter but much more accurate Illumina sequencing reads, whose error rates average <1%. Our hybrid assembly algorithm combines these two types of reads to construct mega-reads, which are both long and accurate, and then assembles the mega-reads using the CABOG assembler, which was designed for long reads. We apply this technique to a large data set of Illumina and PacBio sequences from the species Aegilops tauschii, a large and extremely repetitive plant genome that has resisted previous attempts at assembly. We show that the resulting assembled contigs are far larger than in any previous assembly, with an N50 contig size of 486,807 nucleotides. We compare the contigs to independently produced optical maps to evaluate their large-scale accuracy, and to a set of high-quality bacterial artificial chromosome (BAC)-based assemblies to evaluate base-level accuracy.
- Received July 26, 2016.
- Accepted January 18, 2017.
- Published by Cold Spring Harbor Laboratory Press
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