Taq pro universal sybr qrt pcr master mix

cDNA was generated from mRNA using OligodT primers with a Transcriptor First Strand cDNA Synthesis Kit (HiScript III All-in-one RT SuperMix Perfect for qRT-PCR, Vazyme). An RNase inhibitor was used to prevent degradation. Amplification was performed using Taq Pro Universal SYBR qRT-PCR Master Mix (Vazyme) and specific primer sets (Supplementary Table 3). mRNA expression levels were normalized by to the expression of GAPDH.

Total RNA from human peripheral blood cells was extracted using the TRIzol Reagent. Reverse transcription was performed using random primers and M-MLV RT (Promega) following the manufacturer’s protocol. LncRNAs sequence information comes from the Ensemble database (https://grch37.ensembl.org/index.html), using the national center for biotechnology information (NCBI) online tool to design primers and blasts.
The qRT-PCR reactions were executed using 2 × Taq Pro Universal SYBR qRT-PCR Master Mix (Vazyme) and carried out on a QuantStudio DX (Applied Biosystems Inc., Foster City, USA) according to the manufacturer’s protocol as follows: 95℃ for 30 s, followed by 40 cycles at 95℃ for 10 s, and at 60℃ for 30 s. The relative expression levels of lncRNAs were calculated using the 2^−∆∆Ct method. The above primer sequences are listed in supplementary table S1.