CD8+ T cells activated in vitro within whole splenocyte mixtures
organism strain
C57BL/6J
agent
Nos2-/- FRCs
Sequenced DNA Library
library_strategy
ATAC-seq
library_source
GENOMIC
library_selection
other
library_construction_protocol
Pelleted cells were lysed in 50 μl of reaction mix (25 μl of 2× TD, 2.5 μl of Tn5 enzyme, 0.25 μl of 2% digitonin, 22.25 μl of nuclease-free water) as previously described (Corces et al., Nature Genetics, 2016). Reaction was incubated at 37 °C for 30 min with agitation at 300 RPM. DNA was purified using a QIAgen MinElute Reaction Cleanup kit and Nextera sequencing primers ligated using PCR amplification. Agencourt AMPure XP bead cleanup (Beckman Coulter/Agencourt) was used post-PCR and library quality was verified using a Tapestation machine. Library construction was performed as per standard ATAC-seq protocol, as outlined in Corces et al., Nature Genetics 2016