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RT-qPCR validation

BH Benxia Hu
XL Xin Li
YH Yongxia Huo
YY Yafen Yu
QZ Qiuping Zhang
GC Guijun Chen
YZ Yaping Zhang
NF Nigel W. Fraser
DW Dongdong Wu
JZ Jumin Zhou
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1 μg RNA was reverse transcribed using a primeScript RT and DNA Eraser (TaKaRa, DRR047A) Reagent Kits and stored at −80 °C. Real time PCR was run in triplicate with 50 ng cDNA using FastStart Universal SYBR Green Master (Roche, 04913914001) and ABI7900HT. Relative differences were determined using the ΔΔCt approach. ΔΔCt = (Ctinfection−Ct18S rRNA)−(Ctunfection−Ct18S rRNA). The fold enrichment value is 2−ΔΔCt.

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