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Ambiguously aligned positions and gaps were excluded from alignment, leaving 805 unambiguously aligned positions; the alignment is available from the authors upon request.
The poorly aligned N-termini (~70 80 amino acids for the large subunit and ~40 amino acids for the small subunit) were excluded from alignment.
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Poorly aligned regions (>50% gap in local alignment) and insertion-deletions were excluded from alignments resulting in alignment lengths of 286, 285, 724, 628, 317 and 785 residues for agt, pgm, dbe, be, gbss & ss and pho respectively.
A repetitive region of the control region was excluded from the alignment due to ambiguous alignment and missing data in the NCBI sequences.
Gapped sites and CpG dinucleotide sites were excluded from the alignment blocks, and the alignments in which 80% or more of sites remained were subjected to the subsequent analyses.
Poorly aligned and duplicated sequences were excluded from the alignment.
Sequences aligning poorly with the other CIRs were excluded from the alignment and regions containing large insertions were deleted (Additional file 1).
All sampled sequences were first aligned using MAFFT (E-INS-I, version 6), manually realigned, and then ambiguously aligned sites at the boundary of ORF2 and 3′-UTR were excluded from the alignment.
We manually inspected multiple sequence alignments to identify common sites of the 18S rDNA : large insertions occurring in some sequences were excluded from the alignment to get consistent divergence estimate across pairwise comparions.
LOC_Os03g58990, which has a truncated protein sequence, and LOC_Os04g22080, which includes a retrotransposon-gag sequence in place of Domain B, are excluded from the alignment.
The region of the imperfect 70-bp repeats was excluded from each alignment.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com