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To correct for possible pre-test differences between sequences, we calculated a regression between the pre-test difference (x-variable) and the post-test differences (y-variable) and tested whether the intercept was significantly different from zero.
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To establish if the divergent intensities of signals from solitary LTR probes are a result of differences between LTR loci, or between probes within LTR sequences, we calculated the average signal intensity for each LTR locus.
To determine why the number of unique distances reaches a plateau while the total number of pairwise distances increases quadratically, we need to examine how the distances between sequences are calculated.
The percentage of nucleotide identity between sequences was calculated using DnaSP (Librado and Rozas 2009).
Distances between sequences were calculated by using MEGA version 4 (26 ) with the P-distance analysis option.
Pairwise distances between sequences were calculated using TREE-PUZZLE [ 57] using the WAG [ 55] +Γ+I model.
Percentage identity and biochemical similarity between sequences were calculated using ProtParam http://www.expasy.ch/tools/protparam.html[ 29].
P-distance between sequences was calculated for BuT2 and for the nuclear genes using Mega5.2 (Tamura et al. 2011).
Distances between sequences were calculated using the FastTree program (Price et al. 2010) that produces log-corrected distances calculated with the BLOSUM45 amino acid similarity matrix.
Within each Helitron family, the frequency distribution of the number of pair-wise differences between sequences was calculated with Arlequin v. 3.11.
Distances between sequences were calculated using the JTT matrix, with and without rate variation among sites (the gamma parameter was set to 1.2).
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