GSM8173920: WT_replicating [FL_30]; Saccharomyces cerevisiae; OTHER
Resuspend cells in 500 μl of buffer containg 1.2 M Sorbitol, 200 mM Tris-HCl (pH 8.0), 20 M EDTA, and add 50 μl of 10 mg/ml zymolyase T20 and 0.5 μl β-mercaptoethanol. Incubate samples at 37 °C for 30 min in a water bath. Pellet cells by centrifugation at 16,000g for 2 min at RT, remove the supernatant and set up the following reactions for each sample: 50 mM Tris-HCl (pH 8.0), 100 mM EDTA, 100 mM NaCl, 0.5% SDS, 83.3 mM Sorbitol, 0.33 mg/mL Protease K, 83.3 μg/mL RNaseA. Incubate at 55 °C in a water bath for 2–6 h. Using ethanol precipitation method to pellet the genome DNA.  Resuspend the DNA pellet in TE buffer. More than 1 µg of DNA with good integrity was required. Library preparation and sequencing were commercial services by BGI with a DNBseq platform following the procedure of: DNA fragmentation; fragment size selection; end repair and "A" tailing; adaptor ligation; product size selection; PCR; library QC.