To distinguish AS events, the reverse transcription (RT)-PCR was performed. Briefly, the RNA used for Iso-Seq was isolated using EZ-10 DNAaway RNA Mini-Preps Kit (Sangon, https://www.sangon.com ) and synthesized to complementary DNA using the Hifair® II 1st Strand cDNA Synthesis SuperMix for qPCR gDNA digester plus (Yeasen). Isoform-specific primers were designed using Vector NTI software (Supplementary Table S1 ). PCR amplification was performed using 2 × Taq PCR Master Mix (K1034, https://www.apexbt.com/ ) and the PCR products were showed in agarose gel stained with GoodView™ Nucleic Acid Stain (Apr-13-2021, HGV-II, https://www.sbsbio.com ).
BlazeTaq™ SYBR® Green qPCR Mix 2.0 (GeneCopoeia, USA) was used for RT-qPCR reactions on a CFX96 Real-time System (BIO-RAD, USA). The amplification program was briefly described in previous study [73 (link)]. To ensure the precise and reproducible results, each sample was replicated three times and Ef1α in potato were used for each sample as an endogenous control (Supplementary TableS1 ). The formula F = 2 –ΔΔCt was used to calculate the relative expression levels of selected genes.
BlazeTaq™ SYBR® Green qPCR Mix 2.0 (GeneCopoeia, USA) was used for RT-qPCR reactions on a CFX96 Real-time System (BIO-RAD, USA). The amplification program was briefly described in previous study [73 (link)]. To ensure the precise and reproducible results, each sample was replicated three times and Ef1α in potato were used for each sample as an endogenous control (Supplementary Table