Premix ex taqtm 2 tli rnase h plus kit

To validate the RNA-Seq data, we randomly selected 2 of circRNA and 2 of miRNA for qRT-PCR analysis. Total RNA was extracted from each skin tissue sample, and then reverse-transcribed into cDNA using PrimeScript^TM RT reagent Kit with gDNA Eraser (Takara, Dalian, China) according to the manufacturer’s instruction.
Quantitative PCR (qPCR) was performed using the SYBR^®, Premix Ex TaqTM II (Tli RNase H Plus) Kit with a Bio-Rad CFX Manager 3.1 real-time PCR system (CFX96^TM Real-Time PCR, Bio-Rad, United States). The circRNA expression were normalized to GAPDH as an endogenous reference transcript, the miRNA expression were normalized to that of U6 (Zhong et al., 2019 (link)). The relative expression levels of circRNA and miRNA were calculated using the 2^–ΔΔCt method. The specific primers for each gene are listed in Supplementary Table 2. Data shown represent the means of 3 experiments.

To validate the RNA-Seq data, we randomly selected 3 of circRNA, miRNA and mRNA for qRT-PCR analysis, respectively. Total RNA was extracted from each hippocampal tissue sample, and then reverse-transcribed into cDNA using PrimeScript^TM RT reagent Kit with gDNA Eraser (Takara, Dalian, China) according to the manufacturer’s instruction.
Real-time quantitative PCR (RT-qPCR) was performed using the SYBR^® Premix Ex Taq^TM II (Tli RNase H Plus) Kit with a Bio-Rad CFX Manager 3.1 real-time PCR system (CFX96^TM Real-Time PCR, Bio-Rad, United States). The relative circRNA and mRNA expression levels were calculated using the 2^–ΔΔCt method and were normalized to GAPDH as an endogenous reference transcript. miRNA expression levels were normalized to that of U6. The specific primers for each gene are listed in Supplementary Table 1. Data shown represent the means of three experiments.