Line gene 9600 plus qrt pcr detection system

Total RNA was extracted from the cells using the Trizol (CW0581, CWbio, Beijing, China). One microgram of RNA was used to synthesize cDNA using a reverse transcription reagents Kit (CW0744, CWbio, Beijing, China). The qRT-PCR analysis was then carried out using UltraSYBR (with ROX) on Line Gene 9600 Plus qRT-PCR Detection system (Bioer Technology, Hangzhou, China) in the one-step protocol. Reactions were initiated at 95 °C for 10 min, followed by 45 cycles of 95 °C for 15 s and 60 °C for 60 s. Melting curve analysis was performed to confirm the specificity of primers. The relative mRNA expressions of hCE1, hCE2, BChE, and PON1 mRNA were normalized to GAPDH and calculated using the 2^−ΔΔCt method. The primers used in the study were listed in Table 3.

By using M-MLV Reverse Transcription Kit (Takara, Dalian, China), we collected total RNA for preparing cDNA through reverse transcription. Supplementary Table S1 explains the primers utilized (Sangon Biotechnology, Shanghai, China). miRcute SYBR Green MasterMix and SYBR GreenMaster Mix (Tiangen, Beijing, China) were adopted for qRT-PCR that was carried out using LineGene 9600 Plus qRT-PCR detection system (BIOER., Hangzhou, China). 2^−ΔΔCt approach was used in data calculation relative to the housekeeping gene of sugar beet Actin. The qRT-PCR assay was carried out thrice, and the data were represented as mean ± standard error (SE).