title: GSM4314422: STAT4_NK_WT_IL2_12_6h_rep3; Mus musculus; ChIP-Seq
library_construction_protocol: FastATAC-Seq: FastATAC-Seq was performed according to a published protocol(Corces et al., 2016) RNA-Seq: RNA-seq was performed as described previously with slight modification (Hirahara et al., 2015). Total RNA was prepared from approximately 30-50,000 cells by using TRIzol  following manufacture's protocol (Life Technologies). RNA-Seq: Total RNA was subsequently processed to mRNA-seq library using TruSeq SR mRNA sample prep kit (FC-122-1001, Illumina). RNA-Seq: The purified libraries were multiplexed and captured on an Illumina flow cell for cluster generation. Libraries were sequenced for 50 single read cycles on HiSeq 2000 or 2500 following the manufacturer's protocols. ChIP-seq: Cells were chemically cross linked by 1% formaldehyde. Cell lysates were made by appropriate sonication and protein-DNA complexes were isolated with antibody. ChIP-seq: Libraries were prepared according to Illumina's instructions. Briefly, DNA was end-repaired and the blunt, phosphorylated ends were treated with Taq polymerase and dATP to yield a protruding 3- 'A' base for ligation of NEBNext adapters which have a single 'T' base overhang at the 3' end. After adapter ligation, DNA was PCR amplified with NEBNext index primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel, and quantitated by Qubit (Invitrogen). ChIP-Seq (size fractionation). The purified libraries were multiplexed and captured on an Illumina flow cell for cluster generation. Libraries were sequenced for 50 single read cycles on HiSeq 2000 or 2500 following the manufacturer's protocols. scRNA-seq: Freshly sorted NK cells (13,000 cells each) were encapsulated into droplets, and libraries were prepared using Chromium Single Cell 3' Reagent Kits v2 according to manufacturer's protocol (10X Genomics). Indexed libraries were multiplexed and sequenced on Illumina HiSeq 3000 instruments according to the manufacturer's instructions. scRNA-seq: The generated scRNA-Seq libraries were sequenced using a 50 cycle (paired-end) with a HiSeq 3000 (Illumina).
attributes.tag: GEO Accession
attributes.value: GSM4314422