GSM4058872: H3K4me2_UV3; Caenorhabditis elegans; ChIP-Seq
UV-treated or mock-treated L1 worms were washed off from plates at 24h-post treatment. Crosslink with 1% formaldehyde and suspend in 1ml ChIP buffer (50mM HEPES, 300mM NaCl, 1mM EDTA pH 8.0, 1% Trition X, 0.1% Sodium deoxycholate, 10% glycerol, Protease Inhibitors), then freeze with liquid nitrogen at -80°C. On the next day, worms were homogenized and sonicated to generate worm lysate. Protein concentration was measured and 100 μg of sample was separated as input, and the rest samples was incubated with H3K4me2 or H3 antibody and Dynabeads™  Protein G (Invitrogen) at 4°C over-night. After incubation, samples were washed twice with ChIP buffer, 5min for each, and washed 1 time with ChIP buffer with 1M NaCl for 5 mins, and Washed 1 time with 1ml ChIP buffer with 500mM NaCl for 5 mins, then Washed 1 time with LiCl buffer for 5 mins. Subsequently, elution buffer ( 50mM Tris-HCL Ph 8.0, 10mM EDTA, 1%SDS) was applied and incubated at 4 °C for 1h. Boil the sample at 90°C for 30min with shaking, and check the sonication efficiency by running DNA gels. Afterward, 2 µl Rnase A was added to sample and input, and incubated for 30min at 37°C.  Then 10 µl proteinase K was added and incubated for 2h at 55°C. NaCl was added to samples and input to reach 200mM, and incubated over-night at 65°C with shaking. Experiment was carried out in triplicate. Libraries were prepared according to Illumina's instructions with Cologne Center for Genomics (CCG) modofication.
GEO Accession
GSM4058872