title: Single cell 5' RNA sequencing of Inscripta INSC1005 (S288C like)
design: Yeast cells were grown to O.D. of 1 and were washed twice with 1mL spheroplasting buffer (50mM Sodium Phosphate buffer pH=7.5, 1M Sorbitol, 10mM EDTA, 2mM DTT, 400g/mL nuclease-free DNA/RNA-free BSA; 1000g for 5 min for centrifugation). ~107 cells were resuspended in 100L of spheroplasting buffer + 0.05U/l Zymolase, and were incubated in a flat bottom 96 well plate at 30C for 20 min without shaking. Cells were washed twice with 1mL wash buffer (10mM Tris-HCl pH 8.0, 1M Sorbitol, 400g/mL nuclease-free DNA-RNA free BSA ; 400g for 5 min for centrifugation) and resuspended in wash buffer at 700/l cell concentration. 3 library preparation was performed according to 10X Genomics Chromium Single Cell 3 Reagent Kits v3 user manual (CG000183). 5 library preparation was performed according to Chromium Single Cell V(D)J Reagent Kits v1 (CG000086).