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Isolation by distance was tested within each major mtDNA lineage using a Mantel test of correlation between genetic and geographic distance matrices implemented in the software package IBD [ 42].
The molecular evolutionary history of the Shiva superfamily toxins was reconstructed using Bayesian inference as implemented in MrBayes version 3.2.1 [ 89], using lset rates = invgamma with the prset aamodelpr = mixed command, which enables the program to optimize between the nine different amino acid substitution matrices implemented in MrBayes.
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The coding sequence of the query gene is aligned to the sequence fragments flanking and including polymorphic sites from [35], [36] using the (ends-free) Needleman-Wunsch dynamic programming algorithm and the FULLMAT substitution matrix implemented in EMBOSS.
To identify putative recombination events, third codon-position, ungapped nucleotide sequence alignments from each cluster were created using MEGA and maximum-likelihood trees were created using the HKY+gamma substitution matrix implemented in PAUP* v.4.0 (Sinauer Associates, Inc. Publishers, Sunderland Massachusetts).
For African-Americans and Caucasians single locus genotypic tests of association (additive model) were performed with generalized estimating equations (GEE) using the independence correlation matrix implemented in SAS PROC GENMODD) statistical software version 9.1 (SAS Institute, Cary, NC).
The state-of-the-art method is a heuristic procedure that examines inter-cluster and intra-cluster distance distributions and gives a partition of the set of taxa in a phylogenetic tree, by considering the patristic distance matrix, implemented in a software named CTree [8].
Our previously developed software package consisted of a Monte Carlo simulation module (implemented in C programming language) and a module for constructing the Jacobian matrix (implemented in MATLAB).
In order to construct a scale-free network and identify important modules, we used a weighted adjacency matrix implemented in WGCNA.
Phylogenetic tree was reconstructed using the neighbor-joining method in which distance was estimated by JTT amino acid matrix implemented in MEGA 4.0 program [ 57].
The phylogenetic tree for the insect sHSPs was first reconstructed by Neighbor-Joining (NJ) method in which distance was estimated by JTT amino acid matrix implemented in the program MEGA 4 [ 48].
Otherwise, all sequences for the coding genes were translated to amino acids and aligned based on the Gonnet series matrix implemented in Clustal X [ 53], and subsequently retro-translated to nucleotides.
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