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RT-qPCR

WC Wenbiao Chen
LG Liliangzi Guo
HX Huixuan Xu
YD Yong Dai
JY Jun Yao
LW Lisheng Wang
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Total RNA was extracted from the samples using TRIzol reagent (15596018, Invitrogen, USA). Subsequently, the High-Capacity cDNA Reverse Transcription Kit (4368813, Invitrogen, USA) was employed to reverse transcribe the extracted total RNA into cDNA, utilizing β-actin as the internal reference. For real-time PCR (RT-PCR) experiments, an ABI7500 real-time PCR instrument (Thermo, USA) was utilized in conjunction with the SYBR® Premix Ex TaqTM (Tli RNaseH Plus) kit (RR820A, TaKaRa, Japan). The PCR reaction mixtures underwent amplification using a real-time fluorescence quantitative PCR instrument from ABI, USA. To analyze the final data, the 2-ΔΔCt method was employed. The primer sequences utilized in these experiments were synthesized by Invitrogen and are detailed in Table S3.

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