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In the absence of an objective universal score to compare alignments of different datasets, we chose the 15 largest alignments and visually inspected them for problems.
BLAST hits with extremely low e-values (1e-30 or lower) and high query coverage were then aligned with either MUSCLE (Edgar 2004) or ClustalW (Thompson et al. 2002) to compare alignments and verify the integrity of these hits.
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We compare the results of our alignment method to PEPPeR and OpenMS, and compare alignment accuracy achieved by different versions of our method utilizing various data characteristics.
First, we visually compared alignments for congruence in their ability to align sections of the alignments (especially the inner core) using AltAVisT [54] and the overall sequence identity calculated from each alignment.
The first was the development of the primer pair ENDOITSf and ENDOITSr (Table 1), created by comparing alignments of Atriplex ITS sequences against the ITS sequences of fungal isolates, and identifying potential fungal primers with minimal affinity for Atriplex DNA.
The invariant tryptophan starting the WRKYGQK motif was used as the reference point for comparing alignments.
Consensus ITS-2 sequences, here called genotypes, were constructed manually by comparing alignments obtained using the Clustal W (version 1.83) algorithm [ 21].
This software compares alignments of the sample protein to other proteins with known structural information to determine a probable structure using statistics.
We compared alignments generated by several sequence and structure alignment programs to determine whether existing methods would align residues at a highly conserved position within RIPs (Lebeda and Olson, 1999).
Using these simulated data, we explore the contribution of a variety of factors by comparing alignments that differ with respect to number of sequences, tree topology, alignment length, and gap pattern.
Most importantly, comparing alignments of genomic and transcriptomic reads to the coding sequences, we found no signs of substitution or insertion/deletion RNA editing in E. gracilis mitochondrial transcripts.
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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