title: GSM6923161: CS, replicate 1, scRNAseq; Triticum aestivum; RNA-Seq
library_construction_protocol: Root tips about 1 cm of 40 wheat (CS) seedlings were collected. The wheat (CS) root tips were collected and placed in RNase-free enzyme solution (20mM MES (pH=5.7), 20mM KCl, 8% D-mannitol, 2% Cellulase R-10, 1%Pectinase, 0.4% Macerozyme R-10,0.02M CaCl2, 0.1% BSA), then the mixture was incubated for 4 h in a shaker (room temperature; 90 rpm/min). The protoplasts were filtered twice through a 40um cell strainers, and washed three times with washing solution (20 mM MES (pH=5.7), 20 mM KCl, 0.1% BSA, 0.4 M D-mannitol). After centrifugation, the supernatant was removed, then the adjusted to a concentration of 1000 cells/μl with 8% mannitol solution. RNA libraries  were prepared with Chromium Single cell 3' Reagent v3 Kits according to the manufacturer's protocol.