GSM8001722: Yeast, synHPRT1 integrated on chrXI, rep 2; Saccharomyces cerevisiae; ATAC-seq
~5x106 cells were taken from each culture, pelleted at 3,000 rcf for 5 minutes, washed twice with 500 μL spheroplasting buffer (1.4 M Sorbitol, 40 mM HEPES-KOH pH 7.5, 0.5 mM MgCl2), resuspended in 100 μL spheroplasting buffer with 0.2 U/μL zymolyase (Zymo Research E1004), then incubated for 30 minutes at 30°C on a rotator. Spheroplasts were washed twice with 500 μL spheroplasting buffer then resuspended in 50 μL 1x TD buffer with TDE (Illumina 20034197). Tagmentation was performed for 30 minutes at 37oC on a rotator and DNA was purified using the DNA Clean and Concentrator 5 kit (Zymo Research D4004). PCR was performed as previously described (Buenrostro, J.D., et al., Curr Protoc Mol Biol, 2015) using barcoded primers for 11 total cycles.