Primescript rt master mix kit

Tissues or cells were processed for the extraction of total RNA, following the method provided by the TRIzol kit (Invitrogen, USA). cDNA synthesis was catalyzed by using the PrimeScript™ RT Master Mix kit (Promega, USA). Then, lncRNA ATPT1-AS1 expression was measured by using the ABI PRISM 7700 system using the SYBR-Green PCR Master Mix kit. Reaction parameter settings: denaturation for 60 secs at 95°C, 45 cycles (94°C for 30 secs and 62°C for 45 secs). The relative expression of lncRNA TPT1-AS1 was analyzed by using the 2−△△CT method.

Total RNA was harvested from the serum samples using TRIzol (Takara, Japan) and then used to synthesize cDNA by the stem-loop method using the PrimeScript RT Master Mix Kit (Promega, United States). qRT-PCR was carried out by using an ABI Q3 system (Applied Biosystems, Foster City, CA). The primers used in the study were purchased from Sangya Corporation (Sangya, China) or RiboBio Corporation (Ribo, China). Thereinto, U6 primer was using the Bulge-Loop miRNA qRT-PCR Primer Set (RiboBio, China) according to the manufacturer’s instructions.
U6 and serum miR-484 (Zhou et al., 2015 (link)) expression were used as a stable endogenous control for normalization in cells and serum samples, respectively.