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Nevertheless, a section of the gel encompassing the correct product size (∼359 bp) was identified using marker ladders and excised from each negative control lane.
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The SSRs with the minimal length of 14 bp were identified using WebSat [ 14].
Direct repeats ≥ 20 bp were identified using VMATCH and then divided into distinct families with MATCHCLUSTER by allowing 15% sequence dissimilarity (-erate option set to 15).
Direct repeats longer than 10 bp were identified using the REPFIND program (Betley et al. 2002) and web-server (http://cagt.bu.edu/page/REPFIND_submit, last accessed June 14 , 2013.
ORFs longer than 150 bp were identified using ORFfinder, codon usage Table 4 (http://www.ncbi.nlm.nih.gov/projects/gorf/). ORFs were used to search the non-redundant NCBI database with BLASTP to annotate and find conserved genes.
The GC- and AT-skews, which indicate compositional differences between the two strands, were calculated according to the formulae of Perna and Kocher [ 68] Exact direct repeats longer than 10 bp were identified using the Reputer program package [ 69].
After constructing an index of repeated sequences using MKVTREE with the -dna -pl -allout and -v options, direct repeats ≥ 30 bp were identified using VMATCH (-d and -l options) and then assigned to distinct families with MATCHCLUSTER by allowing 10% sequence dissimilarity (-erate option set to 10).
To determine whether other symbionts are present in the samples, and to search for potentially unintegrated Sulcia-ALF and Nasuia-ALF sequences, Velvet contigs longer than 500 bp were identified using translated Blastx searches against the NCBI nr database and taxonomically binned with MEGAN4 (Huson et al. 2011).
TATA box was identified using the degenerative pattern KAWWW starting from 40 to 20 bp upstream of the TSS [ 41].
The parSc site was identified using the 16-bp consensus sequence (5′-TGTTNCACGTGAAACA-3′) allowing for one base pair mismatching and only one site was found [ 24].
MAP3K7 was identified using GO.
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