GSM9125274: WT Biol Rep 2; Arabidopsis thaliana; RNA-Seq
Once ap1 cal AP1-GR  and drmy1 ap1 cal AP1-GR plants began to bolt, they were induced with 10 μM Dexamethasone in 0.015% Silwet (vol/vol) three times at roughly 24 hour intervals. About 108 hours (4 ½ days) after the first induction, when flower buds had reached stages 2-3, whole cauliflower-like inflorescences were cut and incubated in protoplasting solution, filtered, pelleted, and washed as described in Satterlee et al. 2020 with the following adjustments: cells were incubated in protoplasting solution for 90 minutes, were passed through a 70 micron filter once and a 40 micron filter twice, and 5 μg/mL final concentration of fluorescein diacetate (FDA) was added after the final resuspension in 50-100 µL wash solution to stain live cells for counting. Two biological replicates were protoplasted per genotype using 0.25-0.6 g of tissue per replicate. Library was constructed according to manufacturers protocol (10x Genomics 3' single cell RNAseq pipeline HT kit v3)