Quantitative Gene Expression Analysis in Strawberry

Before RNA extraction, the achenes were stripped from the frozen strawberry fruit, and only the de-seeded flesh was processed for RNA isolation. RNA was isolated from either the deseeded flesh or seedlings by a modified CTAB method. After DNase I treatment, RNAs were used for cDNA synthesis by using the Primerscript RT reagent Kit with gDNA Erase (Takara). The cDNAs were used as templates for quantitative RT-PCR to measure the abundance of a certain transcript. Quantitative RT-PCR was performed using SYBR Premix Ex Tag (Takara) on a Bio-rad iQ5. Primers used are listed in Supplementary Table S6. Results were analyzed by using the ΔΔCT method42 (link) using GAPDH as the control locus43 (link). Three biological and three technical replicates were performed and analyzed.