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. 2010 Feb 18;5(2):e9281. doi: 10.1371/journal.pone.0009281

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Gao et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A real-time RT-PCR using the green fluorescent dye SYBR Green I was applied to determine the relative concentration of BCG mRNA in iCD8⁺DC and iCD8⁻DC. The BCG mRNA in different DC subsets was amplified as described in Materials and Methods. The BCG mRNA level was normalized to GAPDH mRNA in different DC subsets demonstrated as ΔCt. ΔCt = Ct (BCG)–Ct (GAPDH), the threshold cycle of a BCG and the threshold cycle of the corresponding GAPDH in the same sample.