title: GSM1703823: Cflo.minor.scout.TSA.b9.H3K27ac; Camponotus floridanus; ChIP-Seq
library_construction_protocol: RNA extract protocol: A 15µL aliquot representing 15% of cells in a single brain was prepared after light sonication; Total RNA was purified by phenol:chloroform extraction and purification using RNAse-free Agencourt AMPure XP beads (Beckman Coulter, Inc.). 1µL of ERCC RNA Spike-In Mix (Ambion) was added to each 15µL aliquot to total RNA to facilitate transcript-level normalization relative to cell number and to control for technical variation during library preparation. The mRNA fraction of RNA was purified by selection for poly adenylated transcripts using oligo(dT)25 Dynabeads (Invitrogen); cDNA was produced from total RNA using SuperScript III Reverse Transcriptase (Invitrogen), and second strand synthesis was performed using dUTP to preserve strand specificity; Strand-specific RNA-seq libraries were subsequently produced for these dUTP incorporated dsDNA libraries after one round of linear amplification (LinDA) ChIP extract protocol: Chromatin immunoprecipitation (ChIP) was performed on individual scout brains using 45µL of remaining cell material (60e3–75e3 cells) for a subset of the same brains used for RNA-seq according to Simola and Ye, et al, Genome Res 23, 2013; brain tissue was crosslinked with formaldeyde for 5 min.; After mixing C. floridanus (75µg/IP) and H. saltator (1.88µg/IP) chromatin, IPs were performed using 1µg antibody NEB Next Ultra, NEB Multiplex Barcode Oligos
attributes.tag: GEO Accession
attributes.value: GSM1703823