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Sequence assemblies were performed using different assemblers [ 75, 76], which were improved automatically using a number of configuration tools [ 77- 82] and manual inspection.
A number of test assemblies were performed using other assemblers, and a range of parameters was tested within ABySS, and the final, optimal assembly was performed using a k-mer length of 35 and scaffolding with the paired-end data only.
Genome assemblies were performed using the Velvet v1.2.10 [ 41] genome assembler.
In addition, two independent assemblies were performed using reads from each pool.
Base calling and sequence assemblies were performed using the Phred [ 31, 32] and Phrap [ 33], respectively.
Assemblies were performed using two different programs: Newbler (version 2.5 beta) and MIRA (version 2).
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The analysis of the efficiency of assemblies is performed using GMAP and GSNAP v. 2013 11 27.
A multiple alignment of the de novo genome assemblies was performed using progressiveMauve version 2.3.0 [ 63].
Alignment of RNAseq reads against genome assemblies was performed using Tophat and transcript assemblies were determined using Cufflinks (version 2.1.1, [ 103]).
Gaps in the final assembly were closed with GapFiller (Nadalin et al. 2012) and annotation of the final assemblies was performed using PGAAP [National Center for Biotechnology Information (NCBI)].
Sequence editing and contig assembly were performed using Sequencher v4.1.4 (Gene Codes Corporation) or SeqScape v2.5 (Applied Biosystems) and HXB2 as a reference.
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