GSM5563277: G401 MAK683 H3K4me3; Homo sapiens; ChIP-Seq
Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. A total amount of 50 ng DNA per sample was used as input material for the ChIP sample preparations. Sequencing libraries were generated using NEBNext Ultratm DNA Library PrepKit for Illumina(NEB USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. Briefly,the Chip DNA was purified using AMPure XP system (Beckman Coulter Beverly,USA). After adenylation of 3'ends of DNA fragments, the NEB Next Adaptor with hairpin loop structure were ligated to prepare for hybridization. Then electrophoresis was used to select DNA fragments specified in length. 3 uL USER Enzyme(NEB，USA) was used with size-selected,adaptor-ligated DNA at 37C for15 min followed by5 min at 95C before PCR. Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index(X) Primer. At last. PCRproducts were purified(AMPure XP system) and library quality was assessed on the AgilentBioanalyzer 2100 system.