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The supernatant (50 μL) was used to detect catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) activity by using the corresponding assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
Beads were washed once in corresponding assay buffer.
Since the corresponding assay could be routinely and easily run by any microbiology laboratory, typing data from different groups could be merged and compared without the need to exchange live strains.
Spheroids were cultured for at least 24 h and used for the corresponding assay.
The iCycler Software was used to calculate amplification efficiency of each assay and transcript abundance of each unknown sample was extrapolated from corresponding assay standard curves.
General abbreviation of genes selected for this study and corresponding assay ID and accession number was obtained from the Applied Biosystems website and NCBI database.
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These samples were subjected to the corresponding assays in order to determine the maximum additive percentage to be added according to standards.
Subsequently, cells were washed and used for the corresponding assays.
Expressions of these 27 genes were validated in 63 samples using RT-qPCR with corresponding Assay-on-Demand TaqMan® probes (Table 2) and a set of three stably expressed genes as normalizers, selected also from the microarray data.
The corresponding assays monitor the Wnt and the hedgehog pathway.
The average Tt was around 6 minutes faster than the corresponding assays performed without MboI.
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