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The analytical specificity and sensitivity, intra- and inter-assay repeatability and diagnostic characteristics of the kit were determined and compared with virus isolation, which is the gold standard.
Plasma concentrations of glucose (Sopachem, Ochten, the Netherlands), triglycerides (TG), cholesterol (Instruchemie, Delfzijl, the Netherlands), glycerol (Sigma-Aldrich, Houten, the Netherlands) and free fatty acids (Wako Chemicals, Neuss, Germany; HR(2) Kit) were determined following the manufacturers' instructions.
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The precision of the therascreen IDH1/2 kit was determined using a protocol based on the CLSI EP05-A2 [ 25].
The sensitivity of each kit was determined as a minimum rate of CFU providing the positive result in the PCR assay.
Finally, the activity of bound enzyme in all kits was determined with tetramethylbenzidine/hydrogen peroxide substrate.
Dr. Hamedanizadeh explains that distribution of the kits is determined by the size of the affected population and the number of Rural Health Units in the area.
The concentration of RNA isolated with RNeasy kits was determined by measuring the absorbance at 260 nm in a UV-spectrophotometer.
Serum PSA, IL-12 (Bender Med Systems, ELISA kits Vienna, Austria), and IL-10 (R&D systems ELISA kits) levels were determined using ELISA kits as per standard protocol of manufacturers.
The kit components were determined after the PCR amplification conditions were serially optimized.
Briefly, RNA was extracted from all samples with RNeasy Mini RNA isolation kit, and concentrations were determined using a ND-1000 NanoDrop (NanoDrop Technologies, Wilmington, USA).
Cytotoxic (SRB assay) and anti-HIV RT inhibition (RT assay kit, Roche) activities were determined using ELISA.
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