title: GSM1478330: E12.5_H3K4me3_rep2; Mus musculus; ChIP-Seq
library_construction_protocol: Fragments 150-300 bp were size-selected by 2% agarose gel electrophoresis. Recovered DNA was amplified using Phusion DNA polysome (NEB), multiplexing PCR primer 1.0, and one indexed primer. The amplified libraries were purified with Agencourt AMPure XP beads (Beckman Coulter). The libraries were quantitated using the Quant-iT DNA quantitation kit (Invitrogen). The DNA size distribution of the library was measured by an Agilent Bioanalyzer. RNA-seq and ChIP-seq library preparation were performed according to the instruction of NEB NEXT-generation library preparation kit. 2 µg of pooled RNA were used for two rounds of poly(A) mRNA purification by Dynabeads Oligo(dT)25 (Invitrogen). RNA was reverse transcribed by SuperScript III (Invitrogen) and random hexamer primers. After RNaseH treatment and PolI catalyzed 2nd-strand cDNA synthesis, DNA end repair was achieved by End-it kit (Epicnetre). DNA was then A-tailed with Exo- Klenow (NEB), and adaptors were ligated using Quick T4 DNA Ligase (NEB).
attributes.tag: GEO Accession
attributes.value: GSM1478330