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Physico-chemical parameters were analyzed using aliquots from the unfrozen samples.
Three RT reactions were pooled and all qRT-PCR reactions were carried out using aliquots from the pool.
In parallel, the percentage of different sub-complexes within each binding reaction was quantified by electrophoretic mobility shift assays (EMSAs) using aliquots from the same samples.
Using aliquots from the same quenched assay, a Bruker Daltonics Omniflex MALDI-TOF MS was used to determine the CTADOH concentration.
The assay was repeated multiple times using aliquots from the same sample in order to assess the intrasample variability of the RPAC assay.
LC/MS-MS and YES analyses were performed using aliquots from the same sample extracts to eliminate potential bias from sample splits.
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The coefficient of variation (CV) for replicate measurements of urinary 1-OHPG concentration across batches using aliquots from a single quality control pool was 10.6%.
For the farms in Wales and England, it was possible to sample the batch of colostrum powder being used; aliquots from each farm were BLV positive by AGIDT and ELISA.
Third, the Omic analyses benefited from using aliquots of the same tissue, allowing us to better correlate changes between the proteome and transcriptome and observe variations in intermediates and end-products of metabolism.
To better understand the catalytic mechanism of this enzyme, reactions were carried out using different pH conditions and various ions and inhibitors using aliquots of the peak fractions from Mini-S chromatography.
Each experiment was done in triplicate using aliquots of the DNA isolated from each tissue sample, but processed independently.
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