GSM5087715: atxr5/6 (W)_16C_SmartSeq2_S6_11H; Arabidopsis thaliana; RNA-Seq
Fresh nuclei  isolated from two-week-old cotyledons  grown on 1%MS under long day conditions were isolated as follows.  Roughly 50 cotyledons were finely chopped with razor into 50 ul Partec nuclei extraction buffer (Sysmex America, 05-5002), and stained with 400 ul Partec nuclei staining buffer. Samples were filtered once through a 35um nylon mesh (Falcon, 352235) and subjected to fluorescent activated nuclei sorting. Nuclei were sorted from 2C, 4C, 8C and 16C peaks based on DAPI fluorescence. 50 nuclei from each peak were sorted into individual wells of a 96 well plate. Three to four replicates per ploidy/genotype were collected. Negative controls, with no nuclei sorted into a well, were included in every plate and carried through library preparation and sequencing. Libraries were prepared according to Smart-seq V2 protocol at reduced volume and with few modifications as described previously (Picard, C,L et al, bioRxive, 2020).