title: GSM6288268: L1665-Haloferax_volcanii_exponential_ATAC_1pFA_rep3; Haloferax volcanii DS2; ATAC-seq
library_construction_protocol: Cells were fixed by adding 37% formaldehyde (Sigma) at a final concentration of 1% and incubating for 15 minutes at room temperature. Formaldehyde was then quenched using 2.5 M Glycine at a final concentration of 0.25 M. Cells were subsequently centrifuged at 10,000 g for 2 minutes, washed once in 1x PBS, and centrifuged again at 10,000 g for 2 minutes. Transposition was carried out as above for 15 minutes. The reaction was stopped with the addition of 150 uL IP Elution Buffer (1% SDS, 0.1M NaHCO3) and 2 uL Proteinase K (Promega, Cat # MC5005), then incubated at 65C overnight to reverse crosslinks. DNA was isolated by adding and equal volume of 25:24:1 phenol:chloroform:isoamyl solution, vortexing and centrifuging for 3 minutes at 14,000 rpm, then purifying the top aqueous phase using the MinElute PCR Purification Kit, eluting in 10 uL EB buffer. Transposed DNA was isolated using the MinElute PCR Purification Kit (Qiagen Cat# 28004/28006), and PCR amplified for 10 cycles using the usual ATAC-seq settings. Libraries were purified using the MinElute kit, then sequenced on a Illumina NextSeq 550 instrument as 2x36mers.