Illumina MiSeq paired end sequencing of SAMD00119016
YPD cultured cells were cross-linked by 1 percent formaldehyde at 30degree for 30 minutes. After quenching with Glycine, the cell pellets of 21E9 cells were lysed by YTZ-beads. The extracts were recovered with 400 micro-l of lysis buffer 50 mM Hepes-KOH pH 7.5, 140 mM NaCl, 1 mM EDTA, 1 percent Triton-X100, 0.1 percent Na-deoxycholate., and sheared by Covaris S220 to 100bp average. After pelleting, volume of the soluble fraction was adjusted to 1115 micro-l. 50 microl of the extracts were used for INPUT samples. 10 micro-g of anti-HA F-7 mouse antibody was added to 1ml extract and immunoprecipitated with 100 micro-l of ProteinG-dyanabeads. After reverse-crosslinking and DNA purification, sequencing libraries were prepared by KAPA Hyperprep kit and TruseqHT-compatible adaptors. The ligated libraries were amplified by KAPA library amplification kit and purified by AMpureXP double size selection.
UT_IMCB
DRX123814