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The complete sequences of the mitochondrial control region (895 bp) and the ND2 gene (1,047 bp) were aligned using the computer program Clustal W [ 63]; alignments have been further adjusted by eye.
Nucleotide sequences (1995 bp) were aligned using the software ClustalX [67], after removing a polyglutamine stretch [5], which shows length variability in bats and carnivores versus other taxa, and were translated in MEGA 3.1 [68].
Sequences of the 5'-end of the mtDNA CR (ca. 561 bp) were aligned using the computer program ClustalX [ 33].
Fragments in the range 50-490 bp were aligned using ABI PRISM GeneScan 2.1 Analysis Software Applied Biosystemss) and visualized, scored and exported as binary presence/absence matrix using Genographer 1.6.
All non-satellite, non-RepeatMasked high-quality sequences ≥100 bp were aligned using a Burrows-Wheeler Aligner for designed for long reads (bwa-sw) [ 49, 55], to the canFam2.0 assembly to identify all assembled pericentromere-linked assembled chrUn contigs.
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For read pairs in which an alignment was not found for both reads, the reads were trimmed from 100 bp to 75 bp and were aligned using '-n 0' again.
Briefly, trimmed 36 bp reads were aligned using TopHat and FPKM estimates were obtained using Cufflinks (Trapnell et al., 2012) for all features annotated in Ensembl 52.
Identical RADseq tags and those with one or two mismatches (SNPs or 1 2 bp InDels), were aligned using Novoalign software (http://www.novocraft.com/), and the SAM output file was converted to BAM format and sorted using the SAM tools suite [ 34].
For 12 such loci (picked at random, except for avoiding short loci, <300 bp), the sequences were aligned using Clustal Omega (Sievers and Higgins, 2014) and checked for presences of introns by comparison to the bean genome (Schmutz et al., 2014; http://www.phytozome.net/commonbean.php); degenerate primers were then designed to flank the introns using Primaclade (Gadberry et al., 2005; Table 2).
29 bp single end reads were aligned using TopHat version 2.0.9 against the human reference genome (GRCh37.72).
Peak sequences (typically ~500 1000 bp in length) from each promoter subfamily were aligned using ClustalW [ 35] with default parameters.
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