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Many of these misses are due to overstringent cut offs in terms of minimum gene length.
In our analysis, we used a minimum gene length of 110 bp when running Glimmer3 to find missed genes.
In these cases, it is clear that the minimum gene length setting was the primary cause of these genes being missed.
This appears to indicate that in these cases, the use of a high minimum gene length was the primary cause of these annotations' high missed gene rates.
We also found many short missed genes, suggesting that annotators selected a minimum gene length considerably higher than Glimmer's default of 110 bp.
Looking at the shortest annotated gene in these ten annotations reveals the likely use of a high minimum gene length in the process of annotation.
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In fact, for the annotations of two strains of Yersinia pestis, Z176003 and D182038, all 200+ missed genes were under 300 bp, while the minimum annotated gene length in the original annotations was 300 bp.
Proportion of genes from four newly sequenced strains with probe coverage meeting a minimum percentage of the gene length (100%, 90 %, 80) for probes containing at most one SNP.
We adopted a conservative approach to select genes for SNP analyses based on their RNA-seq coverage, requiring 90% minimum read coverage of each gene length, support for moderate expression level (FPKM > 15) and representation in all three strains.
These duplicated blocks overlapped 1007 genes, with a minimum overlap of 50% of the gene length.
Initially hits were selected based on a relatively weak threshold (≥ 20%amino acid identity over the query length); using the minimum gene set criterion, hits to anf/vnfG, and presence of synteny the initial list was refined, yielding the protein sequences listed in Additional file 2: Table S2, Additional file 2: Table S3, Additional file 2: Table S4.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com