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My point here is that life is a lot like the scene described in the opening of A Tale of Two Cities

RNA isolation and quantitative RT-PCR Total RNA from cells was isolated with Trizol reagent (Qiagen GmbH, Hilden, Germany) and RT-PCR was conducted using a quantitative SYBR Green RT-PCR kit (Applied Biosystems, Warrington, UK) was used with a Step One Plus RT-PCR System (Applied Biosystems)