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For each cDNA sample an internal control (beta actin) was also measured by TaqMan probe.
Typical ion chromatograms of sample extracts obtained from a blank sample, an internal standard or from a sample spiked at the 1 ng/ml are shown in supplementary material Fig. S1.
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For the detection of African swine fever virus (ASFV) by polymerase chain reaction (PCR) in clinical samples, an internal control was constructed to identify false negative results in each reaction.
Sampling an internal organ was a good choice for minimizing the risk of contamination, says Bastien Llamas, a geneticist at the University of Adelaide in Australia who studies ancient South American populations.
During the extraction of the samples, an internal standard (dihydroxybenzylamine) was added to allow the subsequent calculation of the exact original concentration avoiding losses during the process.
To normalize variations across samples, an internal standard (IS) solution (100 μg/mL U- 13C-sorbitol, 5 μL) was added to 100 μL supernatant in a 1.5-mL microtube before it was dried by vacuum centrifugation for 2 3 h (4°C).
Amplification of the housekeeping gene β-actin was measured for each sample as an internal PCR control for sample loading and normalization.
A standard 2 μm fluorescent latex bead suspension (BD Biosciences) was added to the sample as an internal reference and Milli-Q water samples were run as negative controls.
For quantitative purpose, β -apo-8'-carotenal was addeacho each sample as an internal standard prior to extraction (10 μg g−1 of freeze-dried sample).
208Po was added to the sample as an internal tracer.
5 µg of methyl nonadecanoate (C19 0, Sigma-Aldrich) was added to each sample as an internal standard.
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