title: GSM6639285: freshly prepared mLN TN cells with low TTS, biol rep1; Mus musculus; ATAC-seq
library_construction_protocol: 5x10^5 cells were suspended in 200 μl Hypotonic buffer (20 mM Tris pH 7.5, 10 mM NaCl, 3 mM ), and incubated for 15 min on ice. Then, 10 μl 10% NP-40 was added to the sample and vortexed for 10 sec at maximum speed. Nuclei were pelleted by centrifugation at 6,000 rpm for 5 min. Nuclei pelets were suspended in 40 μl 1x Tagmentation buffer. Samples were added with 2 μl Tagment DNA Enzyme 1, and incubated for 30 min at 37 oC. Then, samples were cleaned up with FastGene Gel/PCR clean up kit (Nippon Genetics), and eluted with 20 μl elution buffer. 5 μl of the eluted samples were PCR amplified with index 1 (i7) and index 2 (i5) adapter primers, using PCR Primer Cocktail which was attached to Nextera DNA Library Preparation Kit.