Plat_40S
Cells were collected from 5 ml overnight culture and mixed with 200 l breaking buffer (2% (v/v) Triton X-100, 1% (w/v) SDS, 100 mM NaCl, 10 mM Tris-HCl pH8.0, 1mM EDTA pH8.0, following adding 200 l Phenol/Chloroform/Isoamyl alcohol (25:24:1) and 200 l glass beads (Biospec, 11079105). The mixture vortexed at 2000 rpm for 10min. 200 l ddH2O was added to the mixture. After centrifugated at 12,000 rpm for 10min, the top layer was gently pipetted into a new tube and mixed with 1 ml of 100% ethanol, centrifuged at 12,000 rpm, 10min. After drying the DNA, the pellet was resuspended in 0.4 ml sterile water. For DNA sequencing, the DNA sample was treated with 10 l 10mg/ml RNase for 1 hour at 37. Then 4 l of 5M ammonium acetate and 1 ml of 100% ethanol were adding the mixture, followed by centrifugation for 15 min at 12,000 rpm. 100 l sterile water was used to dissolve the genomic DNA for whole genome sequencing.