Instrument: Illumina HiSeq 2500
Strategy: ATAC-seq
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: ATACSeq was performed as described in Buenrostro et al (Nature Methods 2013). FACS-purified cells were spinned down at 1500rpm for 5 min at 4 degree Celsius and washed once with 50ul cold PBS. Cells were then lysed with 50ul lysis buffer (10 mM Tris-HCl, pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630) and the nuclei were immediately spinned down at 1500rpm for 10 min at 4 degree Celsius. The isolated cell nuclei were then incubated with 50ul of transposase reaction (25ul TD buffer, 2.5ul Tn5 enzyme, 22.5ul nuclease-free water)for 30 mins at 37 degree Celcius. Transposed DNA was purified using the QIAgen MinElute PCR Purification Kit following manufacturer's guide. TD buffer and Tn5 enzyme are from the Nextera DNA Sample Preparation kit (Illumina Cat #FC-121-1030). Tranposed DNA was amplified using a low-cycle number PCR protocol and custom primers designed for multiplexing based on Buenrostro et al (Nature Methods 2013). Each sample underwent 14-16 cycles of PCR amplification. Amplified libraries of transposed DNA were purified with the QIAgen MinElute Kit and sequenced on the HiSeq 2000.