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5 g of pre-digested soil samples were spiked with metal standards in triplicate for metal recovery studies as reported earlier [1, 10].
Real-time PCR was then performed on samples and standards in triplicate.
Inter-assay reproducibility (%CV = 1.3-5.1) accuracyrange(range: 96.7-102.3) were determined by assays of the same six standards in triplicate on two days.
In brief, cell culture supernatants and CXCL-10 standards in triplicate were added to precoated 96-well plate and incubated for 2 h at 22 25°C.
Interday and intraday precisions were done by preparing and applying three different concentrations of standards (in triplicate) on the same day and on three different days, respectively.
Along with standards in triplicate (0, 50, 100, 500, 100, 500, 1000, and 2000 mM Hydroxyproline (Sigma Aldrich)) the samples are assessed by a Bio-Tek Synergy HT (Bio-Tek, Winooski VT) plate reader at 558 nm.
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Intra-day and inter-day precisions were done by preparing and applying three different concentrations of standard in triplicate six times a day and similarly on six different days, respectively.
Each experiment included the kit's standards run in triplicate and samples both from the AML patients and healthy controls.
Reproducibility of the assay was assessed based on the Ct values obtained from testing DNA standard in triplicates.
PCR efficiency was determined by obtaining standard curves in triplicate for all primer sets with 4-fold dilutions of a standard batch cDNA.
Experimental values are represented as the mean ± standard deviation in triplicate.
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