GSM4986903: ATAC_NC_D5_1; Homo sapiens; ATAC-seq
Group #1 - Cranial dissection and dissociation using Accumax, Group #2 - Half-head cranial dissection and dissociation using Accumax, Group #3 - culture dissociation using Accutase and counting to normalize cell input. Cells were processed using the OMNI-ATAC-Seq protocol. Frozen cells were quickly thawed at 37℃ and ATAC-RSB buffer was added to a total volume of 1.5mL. Samples were centrifuged at 500rcf for 5 minutes at 4℃ and the supernatant was removed. Next, cells were resuspended in 100µl of ATAC-RSB-LYSIS and kept on ice for 3-5 minutes, depending on input cell type. To stop lysis, 1mL of ATAC-RSB-WASH was added to each sample and they were again centrifuged for 5 minutes at 500rcf. The supernatant was removed, and cells were resuspended in 50µl of OMNI-ATAC Mix (~100nM concentration of Illumina TDE1 enzyme). Cells were then tagmented on a mixing (500rpm) thermoblock at 37℃ for one hour. Tagmented DNA was recovered using a Qiagen MinElute Kit (#28204), with 21µl of elution buffer warmed to 55℃. Library amplification PCR was performed with the NEBNext Ultra II Q5 2X Master Mix (NEB #M0544S) using Nextera primers for 13-15 cycles.