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The genomic shotgun sequence data were assembled with an ABySS [ 37] assembler, and contig ordering was confirmed by the 95,596 paired-end reads obtained from the 8-kb insert library using the Roche/454 pyrosequencing method on a Genome Sequencer FLX system.
Using the de novo transcriptome assembler tool from Trinity [ 35], the normalized data were assembled with a minimum fragment overlap of 35 bp.
The 454 Titanium standard data were assembled with Newbler, version 2.3.
Illumina data were assembled with AbySS [ 24] using default parameters.
The transcriptome data were assembled with Roche GSAssembler (Newbler; version 2.5).
Sequence data were assembled with the GS De Novo Assembler Software (ver. 2.0.01.14, 2.3, and 2.5.3).
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The Illumina data was assembled with the Velvet assembler (version 0.7.55; [ 45]) with a hash length of 61 and the following options; -ins_length 250 -scaffolding no -exp_cov 16 -cov_cutoff 5, to produce an assembly with a final graph with 51765 nodes, n50 of 2413, max 22432, total 37740533, using 73178660/94020728 reads.
The 454 shred data was assembled with the Sanger sequences using parallel Phrap (High Performance Software, LLC).
Illumina sequencing data was assembled with VELVET [81], and the consensus sequences were shredded into 1.5 kb overlapped fake reads and assembled together with the 454 data.
Illumina sequencing data was assembled with VELVET, version 0.7.63 (Zerbino 2008), and the consensus sequences were computationally shredded into 1.5 kb shreds.
Although the data was assembled with programs used by other transcriptome NGS projects [ 16- 20], the assembly itself was revealed to be a major bottleneck.
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