title: GSM7868682: rad51Δ, rep2; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: 2 ml mid-log-phase cells were collected by centrifugation and resuspended in 400 µl TE solution. Add 400 µl acid phenol (CHCl3) to the cells and vortexed vigorously for 10 s. Incubate for 60 min at 65°C, vortexing every 15 min. Then,  place on ice for 5 min and spin at 16,000 × g for 10 min at 4°C. Transfer the supernatant to a new tube , add 400 µl CHCl3 and vortex vigorously for 10 s, and then spin at 16,000 × g for 10 min at 4°C. Transfer the supernatant to a new tube, add a 1/10 volume of 3 M NaAc, pH 5.2, and 2.5 volumes EtOH. Put the tube at  -80°C for 60 min and centrifugate at 16,000 × g for 10 min at 4°C. Wash the pellet twice with 80% EtOH and dry by air. DNase I treatment was performed at 37°C to remove potential gDNA contamination. cDNA  were synthesized by ProtoScript® II Reverse Transcriptase with d(T)23VN primer according to the manufacturer's manual. Synthesized cDNA hybrids were sonicated to 250 bp using ME220 (Covaris, 70 W, 20% Duty factor, 1000 cycles per burst, 130 s, at 4 – 20°C). Denaturation was performed by heating at 95°C for 2 minutes followed by immediately cooling on ice for 2 minutes. Then,the tailing and single-strand ligation method was used for library preparation used as mDRIP-seq Input library construction.