title: GSM3638742: Rbm7_RIP_1; Mus musculus; RIP-Seq
library_construction_protocol: 50 million ES cells over-expressing HA-Zcchc8 and HA-Rbm7 were collected for 0.3% formaldehyde fixation for 10 min. Nuclear pellets were retained after cell lysis for sonication. DNA was sheared between 200-1000bp using Covaris M220 system. 1/50 volume of sheared chromatin was retained as input. Protein A/G magnet beads were incubated with 5ug HA antibody for each reaction. 1/5 volume of sheared chromatin were immunoprecipated with pre-incubated beads overnight. Beads were washed 4 times and eluted (as well as input) following the protocol. Elution samples were cleaned up using RNeasy Mini Kit (QIAGEN). Purified RNA was used for library construction via KAPA RiboErase Kit, 18 cycles amplification during library construction was used for RIP sample to get the final library. For input RNA, 14 cycles amplification was used.
attributes.tag: GEO Accession
attributes.value: GSM3638742