alias: GSM1673882
title: GSM1673882: CD8-Zbtb32KO_CD25lo_ATAC-Seq; Mus musculus; OTHER
library_construction_protocol: 10 million cells for H3Ac and 2 million cells for ZBTB32 and Blimp-1 ChIP-Seq were harvested, crosslinked with 1% formaldehyde, sonicated, and then subjected to chromatin immunoprecipitation with anti-H3Ac, anti-ZBTB32 and anti-Blimp-1 antibodies. 300-500 bp DNA fragments were obtained after reverse-crosslinking. For ATAC-Seq, 50 thousand CD25hi or CD25lo CD44hi CD8+ T cells were isolated at day 5 LCMV-post infection. The H3Ac ChIP DNA fragments were processed for ChIP-Seq with Illumina adapters and Illumina primers (Primer1.1 and Primer2.1). For Blimp-1 and ZBTB32 ChIP-Seq, the ChIP DNA fragments were prepared with a NEBNext Multiplex Oligos for Illumina (Index Primers 1-12) kit (New England Biolabs) according to the manufacturer’s instructions. After adapter ligation, DNAs were PCR-amplified for 17 cycles and library fragments of 300-500 bp were extracted from an agarose gel. For ATAC-Seq, harvested nuclei were subjected to the transposition reaction with Illumina transposase in 1x TD buffer and library fragments were amplified with indexing primers from Nextera kit.
identifiers.namespace: GEO
identifiers.id: GSM1673882