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Here, we analyzed the whole genome polymorphism of 20 D. solani isolates, including the type strain IPO2222T, collected from different geographic locations, dates of isolation and plant hosts.
The dates of isolation of the virus were considered as contemporaneous (isochronous).
However, the samples used in these analyses originated from diverse regions of Africa and the dates of isolation ranged widely.
Further study of temporal population structure made use of regression analyses between tree root-to-haplotype distances and haplotype dates of isolation [63], as implemented by Path-O-Gen v1.1 software (by A. Rambaut; http://tree.bio.ed.ac.uk/software/pathogen).ac.uk/software/pathogen
Furthermore, regression analyses showed that the amount of genetic variation accumulated by haplotypes (i.e. tree root-to-haplotype distances) was significantly correlated with their dates of isolation (Table 3).
The correlation here between tree root-to-haplotype distances and haplotype dates of isolation was statistically significant but not large, which hinted that rate variation might occur within the clonal group.
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All 107 F. tularensis subsp. holarctica isolates showed the same fingerprint pattern by PFGE with the restriction enzyme XhoI, irrespective of their geographic origin, host, or date of isolation (isolate TU41 was not typeable by PFGE analysis).
We used the jModelTest program59 to select the best-fitting substitution model for our sequences according to the Bayesian information criterion, and the date of isolation of each isolate was used to calibrate the tree, and finally these analyses were performed using a log-normal relaxed molecular clock, which allows for different mutation rates on different branches.
Patient information such as age, sex, clinical presentation, isolated site and date of isolation are given in Table 1.
The results indicated that the number of SNPs from the root varied between the different clades and that the root-to-tip distances of individual isolates correlated with their date of isolation (R2 = 0.44; Fig. 2), suggesting that B. pertussis adapted through successive SNP accumulation over time7,21.
MLST showed that, regardless of their geographic origin, date of isolation, or isolation source, chromosomal cpe isolates, i) assemble into one definitive cluster ii) lack pfoA and iii) lack a plasmid related to the cpe plasmid.
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