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Each PCR reaction was repeated three times, and the average median threshold cycle values were used for analysis.
The slope and correlation coefficient of each standard curve were calculated based on the median threshold cycle (CT) values measured for replicates of each dilution point ranging from 106 to 101 DNA templates.
Fold change and median threshold cycle for all cysteine protease genes showing significant difference at P < 0.01 between osteoarthritis (OA) versus fracture to the neck of femur (NOF).
Fold change and median threshold cycle for all metalloprotease genes showing significant difference at P < 0.01 between osteoarthritis (OA) versus fracture to the neck of femur (NOF).
Additional file 2: Median "Threshold Cycle" (Ct) values as obtained for positive and undetermined qPCR tests in fecal samples obtained from the sampled population in the municipality of Barra Mansa/RJ - 2011.
Additional file 1: Median "Threshold Cycle" (Ct) values as obtained for positive and undetermined qPCR tests in feces samples obtained from the sampled population in the municipality of Barra Mansa/RJ - 2011.
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The cutoff value for high vs. low HRG mRNA levels was set at the median delta threshold cycle.
The median low threshold cycle (CT) value for suspensions that generated sufficient DNA for whole genome sequencing (27.3, range 23.5-30.9) wasignificantlylowerwer than for those that did not (30.5, range 28.1-34.0) (p = 0.009).
A biomarker-positive patient was prospectively defined as a patient with HRG mRNA expression above the cutoff, which was specified as the median delta threshold cycle (∆Ct) value in the blinded sample set.
NOF, fracture to the neck of femur; OA, osteoarthritis; Ct, threshold cycle; ND, not detected (median Ct = 40).
Prior to low-density array analysis, samples were assayed for 18S rRNA to ensure that all samples were within 1.5 threshold cycle (Ct) of the median value as a baseline quality control.
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