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To further explore the evolutionary relationships between these four groups of transposases, we used the multiple alignment described above to perform a Bayesian phylogenetic analysis.
The trimmed multiple alignment described above was used to drive a maximum likelihood phylogenetic tree with PhyML [ 42] and the best fitting amino acid substitution model selected with ProtTest [ 43].
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The number of SNPs in each of the subdomains was calculated from the structure-informed multiple sequence alignment described in the previous section.
Specific sites in M13 protein sequences were compared using the multiple sequence alignment described above.
Secondary structure prediction was performed using the last 160 amino acids of the multiple sequence alignment described above.
Similarly, we also elucidated common TFBS among the three topmost conserved human regulatory sequences after multiple alignment (as described below) of NRF2 and NFKB1 sequences.
Furthermore, we took position-specific independent counts (PSIC [ 31]) and adhered to a protocol necessary for sequence extraction and generation of multiple alignment as described elsewhere [ 17].
For the Established Repeat reference data 15 representative orthologs (seven orthologs in case of metazoan-specific proteins) from different taxa were selected from the multiple sequence alignments described above.
Briefly, we sequentially performed gene-wise multiple alignments as described above on all 1729 core genes from the 53 strains used in the study.
The details of multiple alignment construction are described below.
Intron boundaries were mapped onto this alignment and shared positions were defined as introns occurring at exactly the same nucleotide in the multiple alignment essentially as described previously.
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