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Maximum likelihood and minimum evolution phylogenies were inferred from each resulting multiple sequence alignment.
Minimum evolution phylogenies were inferred using fastme [ 39] based on the distance matrix calculated with protdist found in PHYLIP.
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Phylogenic trees were generated using MEGA 3.1, a minimum evolution phylogeny test and 1000 bootstrap replicates.
Genetic distances were obtained from the distance matrix calculated for the minimum evolution phylogeny.
Minimum evolution phylogeny with bootstrapping (500 re-samplings) was performed using MEGA 3.1 [ 34].
Phylogenetic analyses were conducted using MEGA version 3.1 [ 78] using a minimum evolution phylogeny test and 1000 bootstrap replicates.
Parsimonious informative columns of the final alignment were submitted to minimum evolution phylogeny as implemented in the MEGA4 program [ 66].
Phylogeny of MYB proteins were generated from alignments (using Clustal W) and molecular evolutionary analysis conducted using MEGA version 4.0.2 [ 38] (using minimum evolution phylogeny test and 1000 bootstrap replicates).
To estimate the divergence time in different clusters of L. casei, the minimum evolution phylogeny for the 53 strains based on concatenated sequences of the five MLST loci that could be rooted with homologous genes in the closely related species P. pentosaceus (>90% nucleotide sequence identity over a minimum alignment length of 90% of both genes) was used.
The minimum-evolution phylogeny [ 35] was reconstructed as implemented in MEGA4 [ 36] using the JTT amino-acid substitution model [ 37] with 1000 pseudoreplicates for bootstrap analysis.
Phylogenies were estimated by using a minimum-evolution method (11 ), and evolutionary distances were computed by using the Tamura-Nei method (12 ).
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