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The root tip region (0.5 cm in length from root tip) were harvested and digested for 2 hours at room temperature in the RNase-free enzyme solution (1.5% cellulase R10, 1.5% macerozyme R10, 0.4 M mannitol, 10 mM KCl, 10 mM CaCl2 and 0.1% BSA). The protoplasts were filtered with a cell strainer (40 in μm diameter, FALCON, 352340), concentrated and washed 3-4 times with 8% mannitol at room temperature. The protoplast viability is determined by trypan blue staining. The ratio of viable cell should be more than 85%. The concentration of protoplasts was adjusted to 1500 ~ 2000 cells/μL. The cells were then processed with the 10xGenomicsZhang et al: Arabidopsis root atlas Single Cell Protocol (CG00052, RevC) |
RNA-Seq |
TRANSCRIPTOMIC |
PCR |
PAIRED
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