title: GSM7901712: Eomes-KO CD4 T cells (replicat 2) [ATAC]; Mus musculus; ATAC-seq
library_name: GSM7901712
library_construction_protocol: ATAC-seq was performed as previously described (Adoue et al., 2019) with minor modifications. Briefly, 50 000 cells were lysed in ice-cold lysis buffer and the transposition reaction was performed using the Tn5 transposase at 37°C for 30 min. DNA was purified using the QIAGEN MinElute kit (QIAGEN). The libraries were prepared using Tagment DNA Enzyme and Buffer kits (Illumina), NEBNext High-Fidelity 2X PCR Master Mix (NEB, catalog # M0541S) with custom sequencing primers (Ackermann et al., 2016; Buenrostro et al., 2016, 2013; Tsompana and Buck, 2014). The libraries were purified twice using AMPure XP beads (Beckman) following a double-sided protocol to remove primer dimers and large fragments. Libraries quality was assessed using the NGS High Sensitivity Kit on the Fragment Analyzer (Agilent Technologies, Santa Clara, USA).