title: ATAC-seq WA09, Day 21, Replicate 1
design: ATAC-seq was performed on 50.000 to 100.000 cells according to the manufacturers instructions (Active Motif, #53150). The cells were pelleted and washed with 100 L of ice-cold PBS. They were then lysed with 100 L of ATAC Lysis Buffer on ice for 3 minutes and centrifuged at 500 g for 10 minutes at 4C. The nuclei were incubated in 50 L of Tagmentation Master Mix at 37C for 30 minutes in a thermomixer set to 800 RPM. During this process, the Tn5 Transposase probed accessible DNA regions in the nuclei by inserting sequencing adapters into open regions. The tagged DNA was then purified and PCR-amplified. Prior sequencing, the libraries were quantified by qPCR using the KAPA library quantification kit for Illumina platforms (Roche) using QuantStudioTM 12K Flex Real-Time PCR System (Thermo Fisher Scientific). All samples were normalized according to concentration and pooled. Libraries were loaded and sequenced using the NovaSeq 6000 System (Illumina).