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The concentration of the templates was adjusted by amplification using primers for Dd-Ig7 (mitochondrial large ribosomal RNA) or As-elp3 (a subunit of the RNA polymerase II elongator complex).
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All DNA templates were adjusted to a concentration of 50 ng/μl with distilled water for PCR tests.
Given that ticks were not homogeneously engorged and DNA concentrations differed among tick extracts, DNA templates were adjusted to a stock concentration of 40 50 ng/μL.
All single-primer templates were adjusted to the same concentration of 0.001 ng/μL prior to the single-primer amplification test.
Their stomachs were prepared as wholemounts and processed with tetramethylbenzidine. Inventories were obtained with a counting grid that was systematically positioned throughout the wholemounts by the use of a sampling template that was adjusted for stomach size and shape.
The translation reaction mixture contained 70% (v/v) of the supplemented LTE, 10% (v/v) of OE-PCR mixture or 10 nM of the linearized plasmid as a template and was adjusted to the final volume with nuclease-free mQ water (Ambion).
Template input was adjusted to obtain a cluster density of 700 900 K/mm2.
The amount of template from each sample was adjusted until PCR yielded equal intensities of amplification for β-actin.
The concentration of pBS sequence template, either cut or intact, was adjusted to be about 0.1 nM and to be equal in the two samples.
The amount of cDNA template used for the RT-PCR was adjusted on the basis of amplification with primers specific for human GAPDH: forward 5′-CCTCCAAAATCAAGTGGGGCG-3′ and reverse 5′-ACCACCAGGTGCTCAGTGTAG-3′.
The quantity of input DNA templates for PCR from each environmental sample was adjusted based on the relative 16S rDNA PCR result.
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