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Inspecting the sequence of the cab12 promoter for the ACGT core sequence of bZIP factor binding sites reveals 12 positions for this motif (data not shown).
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Since the RefSeq annotations of chicken and zebrafish are incomplete, we identified additional orthologous exon pairs by projecting the human annotation on the other species' genome and confirming the orthology of the projected exons by inspecting the sequence alignment of the projected exons in the representative species.
We focused on known NF-κB target genes and inspected the sequences for the presence of κB sites from –10 kB to +1 kB of the transcription start site (a subset of these sites are listed in Figure 2A).
To test the reliability of these called differences, we inspected the sequence reads of 307 selected genes (exhibiting in total 647 non-synonymous variants) using the Integrative Genomics Viewer (IGV) [ 42].
We have manually inspected the sequence alignment of the homologous flanking ±100-bp regions.
We therefore inspected the sequence alignments of NA12716 and NA12760 in this region (SNVs 227 to 277; additional file 17).
Looking at the highly thermostable oxidase HotAldO for inspiration, we inspected the sequence alignment of AldO and HotAldO (Fig. 1) and identified the HotAldO residues that corresponded to the AldO residues with high B-factors.
We inspected the sequencing reads of this variant to test whether the deleterious allele was skewed towards homozygosity and found that of the sequencing reads overlapping this variant position, 46 supported the alternate alleles, whereas only 28 supported the reference allele, suggesting that the alternate allele is homozygous in the mosaic cell line.
To identify putative cis-elements present in the NFS1 and NFS2 promoters, we inspected the sequences 1,500 bp upstream of the transcriptional start site of all genes using the Plant Cis-Acting Regulatory Elements (PlantCARE) database [ 28].
By inspecting the RVD sequences of PthA2 and PthA4, we presumed that they would target common host genes.
Finally, Manser et al. had to visually inspect the primary sequence of Syx to uncover its RBD.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com