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Linearity and efficiency of the protocols were assessed using a standard curve constructed by the amplification of plasmid standard pools with a total number of 5·109 copies/ml including different mutant DNA frequencies, ranging from 100% to 0.39%.
For the estimation of mutant DNA frequencies standard curves were produced by plotting the threshold cycle values (Ct values) versus the logarithm of % frequency of mutant (V136) DNA in the standard plasmid DNA pools.
The ability of the basic real-time LCR protocol to accurately quantify mutant DNA was assessed using genomic DNA pools that were artificially prepared to contain different mutant DNA frequencies (50%, 25%, 6.25%, 1.56%, and 0.39%).
DNA pools with a total number of 5·109 copies/ml, defined as mutant DNA frequencies of 100 %, 25, 6.25%, 1.56.25nd 0.39%, were generated by serial dilutions of mutant V1.56plasmid DNA in 1∶4 ratios with the wild-type A136 plasmid DNA.
Values of ΦST, basedon mitochondrial DNA frequencies and haplotype divergences, betweensamples of North Pacific red king crab.
There was more variation between observed and expected DNA frequencies as the minor allele frequency increased in all datasets, showing that allele frequency estimation may be more accurate at loci with lower minor allele frequencies.
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DNA frequency histograms were obtained using FlowJo software (Treestar, Oregon, USA) using the Dean-Jett-Fox model.
Splinter et al. (2006) [33] have recently shown that in CTCF null cells, there were reduced DNA-DNA interaction frequencies between the sites that normally bind CTCF in the mouse β-globin gene locus.
The relationship between DNA uptake frequencies versus selection is examined and the implications for the open release of genetically engineered microbes is discussed.
Low interspecies DNA transfer frequencies suggested substantial divergence [11].
Mitochondrial DNA haplogroup frequencies were calculated by counting from the observed genotypes.
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