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One of the stocks will then be used for normalising the DNA concentrations in 96 deep well (DW) plates.
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The amyloid beta (A4) precursor protein was used for normalising the cDNA concentration of each sample.
A dilution series (1 4 to about 1 10,000) of a reference cDNA pool (mixture of cDNA preparations of RNA derived from six different cell lines) was used for normalising gene expression.
Another approach that can be used for this purpose is to normalise parameters to control and consider the percentage change in an attempt to differentiate direct and downstream mechanisms [ 20, 21].
Because different exposure times have to be used for the holographic and for the conventional microscope, to compare the results I normalised the two measurements.
SyBr green semi-quantitative assay was then completed on the biological samples and on three technical replicates on a StepOne Real time system (AB Applied Biosystem) and the ∆∆ method [ 62] was used for quantifying the relative expression levels normalised against the actin gene.
The normalised intensity values were used for independent pairwise comparisons for each tissue, using the empirical Bayes approach implemented in limma by assigning the samples to two groups according to genotype (40,XX and 39,XO).
Mean incidence and peak incidence were log-transformed to normalise the data; a logit transformation was used for outbreak size.
To normalise the expression data, a single control gene, HvGAPdH, was used for this single tissue, single time point experiment.
All plasma AUCs were normalised for their matching control group, and these relative plasma AUCs were used for comparison.
Filtering can thus be used to normalise their spectra for clearer data analysis as shown in Figure 16.
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