the total RNA from each sample was isolated using the SteadyPure Universal RNA Extraction Kit
Label
SYBR Green
Label protocol
RT-qPCR amplification was performed in the Applied Biosystems Stepone system (Thermo, USA). The real-time PCR volume of 10 µL contained 5 µL 2X SYBR® Green Pro Taq HS Premix (Accurate Biology, China), 0.5 µL primer F (5 µmol·L−1), 0.5 µL primer R (5 µmol·L−1), 1.5 µL cDNA, 2.3 µL ddH2O, and 0.2 µL Reference Dye. RT-qPCR conditions were set as follows: 95 °C for 30 s, 40 cycles (95 °C for 5 s, 60 °C for 30 s), and an infinite hold at 10 °C. The melting curves ranging from 60 °C to 95 °C were evaluated in each reaction to confirm the specificity of amplified products.
Hybridization protocol
n/a
Scan protocol
n/a
Data processing
For the normalization it uses the most stable reference genes For the normalization it uses combinations of the most stable reference genes For the normalization it uses the least stable reference genes Matrix normalized worksheet reports normalized signal (against housekeeping genes)
