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The adult livers were clustered independently of the embryonic tissues, when each duplicate of samples taken from the same adult was clustered together and seemed nearly identical, confirming the reliability of the microarray results.
In brief, duplicate of samples and LIF standards (serially diluted in phosphate buffer) were added to 96 well plate pre-coated with mouse-anti human LIF mAb.
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For statistical analysis, duplicates of samples (e.g., abdominal swab and abdominal liquid/syringe) were removed.
Case and control DNA were genotyped together on the same plates with duplicates of samples (10%) to assess intraplate and interplate genotype quality.
Case and control DNA was genotyped together on the same plates with duplicates of samples (15%) to assess intraplate and interplate genotype quality.
Values are average from duplicate groups of samples.
Furthermore, method performance was carefully monitored using multiple internal standards (5 to 10 depending on method, including analogues, 2H and 13C labeled metabolites) and duplicate analysis of samples.
The coefficients of variation of duplicate analysis of samples were between 5% and 36%.
The relative standard deviations of duplicate analyses of samples were within 5%.
Duplicates of sample cDNA were then amplified on the 7900HT Fast Real-Time PCR System (Applied Biosystems) using the Fast SYBR Green RT-PCR kit (Applied Biosystems) in 96-wells plates (micro amp optical, Applied Biosystems).
Intra assay CV was calculated as the mean of the CV of duplicate samples of synovial fluid and also plotted as a precision profile.
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