Mirna primescript rt reagent kit

Following the manufacturer's instruction, total RNAs were extracted from tissues or cultured cells using RNAiso Plus reagent (TaKaRa, Japan), and roughly quali ed by using 1.5% agarose gels electrophoresis and NanoDrop 2000c Spectrophotometer.
Then the quali ed RNAs (~1 mg) were reversely transcribed into cDNA by using the PrimeScript™ RT reagent Kit with gDNA Eraser or miRNA PrimeScript RT reagent Kit (Takara, Japan) separately for mRNA or miRNA assay. Then according to the manufacturer's guide, RNA levels of target genes in these cDNA were accurately measured by using real-time PCR (RT-qPCR) in the Bio-Rad CFX96 system (Bio-Rad, USA) with SYBR Premix Ex TaqTM II (Takara, Japan). Each treatment performed at least triply three independent times, and each sample triplicates in qPCR. Moreover, to enhance the accuracy, three housekeeping genes in goat ACTB (Actin Beta), SDHA (Succinate Dehydrogenase Complex Flavoprotein Subunit A), and PGK1(Phosphoglycerate Kinase 1)) and mouse (ACTB, GAPDH, and Hprt (Hypoxanthine Phosphoribosyltransferase 1)) were used as an internal control. The 2 -△△Ct or 2 -△Ct methods were employed to calculate the relative RNA levels of target genes. The detailed information for primers was listed in Supplementary Table 1.