GSM4201674: ATAC-seq from Luminal2/BMP k.o CAF; Homo sapiens; ATAC-seq
We lysed 50,000 cells with 50 μl of Lysis buffer (10 mM Tris-Cl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0.1% Tween-20, and 0.01% Triton X-100) and incubated them for 5 min on ice . Following centrifuging at 1,000 g for 10 min at 4 °C, we resuspended the nuclei pellets with 50 μl of transposition mix (25 μl 2X TD buffer (20 mM Tris-HCl pH 7.6, 10 mM MgCl2, and 20% Dimethyl Formamide in sterile water) and 2.5 μl transposase (100 nM final), 16.5 μl PBS, 0.01% Triton X-100, 0.1% Tween-20, and 5 μl H2O) and incubated the mixture with shaking for 1 h at 55 °C in a thermomixer. We purified transposed DNA using DNA Clean and the Concentrator-5 Kit (Zymo Research). We amplified the transposed DNA using the NEBNext® High-Fidelity 2X PCR Master Mix for 10 cycles. We then performed ATAC-Seq using the Illumina HiSeq 2500 platform.
GEO Accession
GSM4201674