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Real-time PCR was performed in parallel in 25 ΞΌl reactions containing 12.5 ΞΌl 2 Γ QuantiTect SYBR Green PCR master mix (Qiagen, Venlo, The Netherlands) and 2.5 ΞΌl 10 Γ QuantiTect Primer Assay (Qiagen)