GSM2845177: ATACseq_Epiboly_Citrine_1; Danio rerio; ATAC-seq
FACS sorted cells were washed with PBS and stored at -80C in lysis buffer. RNA was extracted using Ambion RNAqueous Micro Total RNA isolation kit (AM1931), checked on Bioanalyser, samples with RIN >7 were used to prepare cDNA using Takara Clontech SmartSeq2 V4 kit (634889). Sequencing libraries were prepared using Illumina Nextera XT library preparation kit (FC-131-1024). For ATAC-seq, FACS sorted cells were lysed (10mM Tris-HCl, pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% Igepal) and tagmented using Nextera DNA kit (Illumina FC-121-1030 ) for 30 minutes at 37C. Tagmented DNA was ampli ed using NEB Next High-Fidelity 2X PCR Master Mix for 11 cycles. Tagmentation e fficiency was assessed using Agilent Tapestation. ATAC-Rx was carried as previously described (Orlando et al., 2014), and as described above with the addition of 50% extra Drosophila S2 cells. ATAC-seq libraries were sequenced using paired-end 40 bp run using Illumina NextSeq500 platform.