GSM8640702: ATAC of Mmmyco strain (gDNA), rep1; Saccharomyces cerevisiae; ATAC-seq
Around 1-5 × 106 cells were taken from each culture, pelleted at 3,000g for 5 min, washed twice with spheroplasting buffer (1 M sorbitol, 40 mM HEPES-KOH pH 7.5, 10 mM MgCl2), resuspended in 200 μl spheroplasting buffer with 10 µL of Zymolyase (10 mg/mL), then incubated for 30 min at 37 °C. Spheroplasts were washed twice with 500 μl spheroplasting buffer then resuspended in 50 μl 1× TD buffer with 2.5 µL of TDE1 (Illumina 20034197). Tagmentation was performed for 30 min at 37 °C, 800 rpm with a thermomixer, and DNA was purified using the DNA Clean and Concentrator 5 kit (Zymo Research D4004). PCR was performed using Phusion Master Mix, 11 total cycles. Paired-end sequencing on an Illumina NextSeq2000 (2 x 50 bp) was performed.