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Quality controls were run every day before analysis of samples.
After washing twice with cold medium phosphate buffered saline (PBS), cells were incubated with Annexin V-FITC in dark for 5 min, and then PI was added before analysis of samples with flow cytometry.
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A preliminary assessment of candidate gene expression in the cell lines under investigation following exposure to PBMC was made in pooled cDNA samples before analysis of individual samples.
The solitary case reported as negative by HR-HPV RNAscope demonstrated an expression level for both E6 and E7 that was low by comparison with other samples; however, it was not the lowest and remained within the threshold for detection set before analysis of the samples.
In order to minimize the interference effect, the synthesized polymer was successfully used as a solid phase sorbent for salbutamol extraction, before analysis of real samples by the developed sensor.
Just before analysis of the samples by FACS, propidium iodide (PI) (Sigma, St . Louis MO) was added.
Before analysis of patient samples, the method was validated with respect to accuracy and precision.
Before analysis of the samples by FACS, propidium iodide (PI) (Sigma-Aldrich GmbH) was added (5 μg/mL).
Before analysis of the samples, the sensitivity of the PCR set up was determined by performing the PCR on dilution series of the DNA vaccine spiked with genomic DNA of muscle tissue.
All samples (except samples stained for NOTCH1) were treated for 15 min with 0.1% Triton X-100 and blocked with 2% bovine serum albumin/Triton X-100 in PBS for 15 min at RT. Before analysis of NOTCH1, samples were digested with 8000 units/mL hyaluronidase (Sigma-Aldrich) for 1 hour in 37°C and then digested with 0.05 units/mL chondroitinase AC (Sigma-Aldrich) for 1 hour in 37°C.
Before analysis of the serum samples the serum proteins were precipitated by adding an equal volume of MeCN to the samples.
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