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A serial dilution standard curve was prepared from a pool of all the samples and real-time PCR was carried out.
Total RNA was prepared from a pool of 20 wild type and 20 EP-704/EP-704 3rd instar larvae using the TRIzol reagent RNA protocol following GibcoBRL instructions.
For sample A, protein extracts were prepared from a pool of 40 male antennae (20 specimens) and from a pool of 80 female antennae (40 specimens), being female antennae smaller and less plumose than males.
Each soluble intestinal contents sample was prepared from a pool of 20 midguts from blood unfed A. aegypti females with at least 7 days old, or a pool of 6 anterior midguts from each triatomine species obtained from starving fourth instar nymphs with 10 to 20 days after moult.
Total RNA was prepared from a pool of T2 transgenic plants (n > 50 for each line).
For each time point, RNA samples were prepared from a pool of ~30 plants per replicate.
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The QA/QC materials were prepared from a plasma pool obtained from multiple anonymous pregnant women donors in analysis with standard, reagent blank, and unknown samples.
Genes were cloned by PCR amplification using sequence-specific primers and cDNA prepared from a mixed pool of HH3 to HH6 chicken embryos.
Low-concentration (QCL, 2 5 ng/mL) and high-concentration (QCH, 10 50 ng/mL) QC materials were prepared from a base urine pool obtained from multiple anonymous donors (Ye et al. 2006).
QC materials with low concentrations (QCL) and high concentrations (QCH) were prepared from a base urine pool, obtained from multiple anonymous donors as described previously (Silva et al. 2004), dispensed in 5-mL aliquots, and stored at –20°C.
QC materials of low concentration (QCL) and high concentration (QCH) were prepared from a base urine pool, obtained from multiple anonymous donors, dispensed in 5-mL aliquots, and stored at −20°C.
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