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Plates were sealed in plastic bags and stored at 4°C until the assay was started.
Drug-free human urine samples were obtained from healthy volunteers who gave their informed consent, filtered through a filter paper, and stored frozen until the assay was carried out.
The homohopane index decreased in the first 60 days and increased from then on until the assay was completed (180 days), with an unusual homohopane biodegradation preference C33 > C32 > C31 > C34 > C35 Munoz et al. (1997) showed 25% depletion of C35 pentakishomohopane and only 3% depletion of C31 homohopanes in soils exposed to an oil spill for eight years.
Samples were suspended in 50 µL ultra pure water, and stored at −20°C until the assay was performed.
After 24 hours incubation, plasma aliquots were harvested, and frozen at 70°C, until the assay was performed.
Plasma was isolated from whole blood by centrifugation at 1,000×g for 10 minutes at room temperature and stored at −80°C until the assay was performed.
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For hsCRP, sCD40L and prolactin, venous blood samples were centrifuged within 15 min at 3,000 rpm for 10 min, and the supernatant serum samples were transferred into polypropylene tubes at −80°C until the assays were performed.
Total RNA was extracted from specimens at 7, 15 and 30 days using TRI Reagent (Sigma, Milan, Italy), following the manufacturer's instructions, treated with DNase (Promega) to exclude DNA contamination and stored at −80°C until the assays.
All the sera were stored at -20°C until the assays.
Serum samples were stored at -30°C until the assays.
The serum remained stored at −80°C until the assays were performed.
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