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On the other hand, novel mass peaks only observed in the mixed sample were discovered and may serve as biomarkers for mixtures or contaminations [83, 86].
Heavy and light samples were then mixed 1 1, and 4 μg of each mixed sample were in-solution reduced, alkylated and digested with trypsin.
DNA samples from each mixed sample were also analyzed in order to correct for both allele-specific amplification error and differences in cell number between the individuals extracted from each parental line [ 15].
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Then, 50 g of the mixed sample was transferred into the enrichment medium.
The basic principle of the approach is that a fluorescence pattern of a mixed sample is a superposition of the fluorescent patterns for each target.
The mixed sample was centrifuged (10,000×g at 4°C) for 5 min to obtain the RNA pellet.
A volume of 25 µl (corresponding to ∼13 µl of plasma) of the mixed sample was then transferred onto filter paper and left to dry overnight.
The 10 μL mixed sample was used for 12% SDS-PAGE loading.
Five minutes later, the absorbance at 545 nm of the final mixed sample was measured.
This mixed sample was applied via anodic cup loading on one gel strip.
Fifty microliters of this mixed sample was then applied to an HPLC system (Waters, MA, USA).
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