title: ChIP-seq of human:LHCN-M2 Cells-forH3K36me2 input
design: The cells were corsslinked with with 1% formaldehyde for 10 min at room temperature and quenched by 125 mM glycine. The crosslinked cells were then lysed in lysis buffer (50 mM HEPES-KOH, pH 7.9, 140 mM NaCl, 1 mM EDTA, 10% glycerol, 0.5% NP40, 0.25% Triton X-100, supplemented with cOmplete protease inhibitors (Roche)) and incubated on ice for 10 min. The cell pellets were then washed with wash buffer (10 mmol/L Tris-Cl, pH 8.1, 200 mM NaCl, 1 mM EDTA, pH 8.0, 0.5 mmol/L EGTA, pH 8.0, supplemented with cOmplete proteinase inhibitors) and shearing buffer (0.1% SDS, 1 mM EDTA, 10 mM Tris-Cl, pH 8.1, supplemented with cOmplete proteinase inhibitors). Then, the cell pellet were resuspended in shearing buffer and sonicated with Covaris E220 Evo sonicator to shear DNA from 300 to 500 bp size fragments. Then Purified DNA was used for performing ChIP-seq library preparation by TruSeq ChIP Sample Preparation Kit (Illumina, IP-202-1012, IP-202-1024) according to the manufacturers instructions. Briefly, 5 ng of DNA was used as starting material for input and IP samples. Libraries were amplified using 14 cycles on the thermocycler. Libraries were validated using the Agilent High Sensitivity DNA Kit.  Sequencing was performed with an Illumina HiSeq  with 150-bp paired-end read runs.