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The size and morphology of the samples were inspected using a field emission transmission electron microscope (FE-TEM) equipped with an energy-dispersive X-ray spectrometer (EDX) (FE-TEM, JEM-2100F, JEOL, Akishima-shi, Japan) by operating at an accelerating voltage of 200 kV.
After washing with a saline solution for three times 20 min each, the samples were inspected, using LCSM.
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The sample is inspected using optical microscopy in order to identify thin flakes (<10 nm) lying across the aluminum gate, e.g., a flake as in location (I .. Thin flakes were found to exhibit a characteristic yellowish color on the gate region (Al2O3/Al) with a strong contrast to thicker flakes of ~15 25 nm, which appear distinctly blue-colored (cf. locations (II).
The integrity of the DNA sample was inspected using 1 % agarose gel electrophoresis while its purity was measured at A260/A280 and A260/A230 ratio, respectively, by using a spectrophotometer (Nanodrop 2000, Thermo Scientific).
Mean, standard error, highest posterior density intervals (95%HPD) and effective sample size of likelihood, evolutionary rates and the TMRCA of Carabus were inspected using Tracer 1.5.
Convergence and mixing of the chains were inspected using Tracer version 1.5; all continuous parameters yielded effective sample sizes greater than 200.
Chromatograph profiles were inspected using Chromas 2 software.
Clusters were inspected using the Genotyping Module within Genome Studio.
The capture efficiency was inspected using a microscope to remove samples from the analysis with more than one cell captured.
Convergence to the stationary distribution of the single chains was inspected using a minimum threshold for the effective sample size.
Convergence of the MCMC chains was inspected using TRACER 1.5 [ 75] by visually checking that effective sample size (ESS) for all relevant parameters were well above 200.
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