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An aliquot of the diluted mixed culture was simultaneously plated onto selective and non-selective media for cell counts.
Figure 1 and Figure 2 demonstrate standard curves for SRY and β-globin genes plotting the threshold cycle (CT) against known concentrations of serially diluted mixed DNA.
For identification and calibration of the analytes, ClinCal® serum calibrators 15013 and 15213 (Recipe, Germany) were diluted, mixed together, spiked with internal standard, and prepared as waste water samples to get a 2-point calibration with preconcentration factors of 0.6 and 0.3 for 15013 and 0.4 and 0.2 for 15213.
The PCR products were diluted, mixed with the internal size standard LIZ500 (Applied Biosystems) and loaded on an ABI 3130 Genetic Analyzer.
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cDNAs were diluted ×9, mixed with primers (0.2 mM) and Thermo start master mix (ABgene, Surrey, UK).
Additionally, one-step real-time RT-PCR was performed to estimate the detection limit on c-kit+ cells sorted from bone marrow cells and serial diluted and mixed with c-kit bone marrow cells or with purified myocyte preparation.
The cells were then immediately transferred into Hungate tubes with 5 ml fresh RCM medium, incubated at 45°C for 5 h, diluted and mixed with melted warm RCM-agar medium supplemented with 250 µg/ml kanamycin, and then plated onto Petri dishes inside an anaerobic glove box.
Samples were thawed, diluted, and mixed in phosphate-buffered saline (PBS) immediately before use.
For estimation of platelets activation, 250 μl EDTA blood was diluted and mixed 1 1 in Hepes-buffer.
These stock solutions are then further diluted and mixed with 75% poly ethylene glycol, MW = 600 D) (PEG) for a final PEG concentration of 50% (v/v).
However, our subjects included both women and men so that some underlying amygdaloid contribution may have been diluted by mixed lateralization components in women and men [ 30, 31].
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