GSM5642622: D2_MyoD_2 (Omni-ATAC-Seq); Mus musculus; ATAC-seq
Cultured cells were lysed on ice by cold lysis buffer containing 0.1% NP-40, 0.1% Tween-20, 0.01% Digitonin, 10 mM Tris-HCl (pH 7.5), 10 mM NaCl, 3 mM MgCl2 in Nuclease-free water. Nuclei were collected by centrifugation and subjected then to transposition reaction in 1x Tagment DNA buffer containing 5% Tn5 Transposase (Illumina, 20034197), 0.1% Tween-20, 0.01% Digitonin at 37°C for 30 mins on thermomixer, followed by DNA purification using MinElute PCR purification kit (Qiagen, 28004) Index PCR amplification was performed by mixing 10 µL of transposed DNA with 10 µL nuclease-free water, 25 µL NEBNext High-Fidelity 2X PCR Master Mix, 2.5uL of each 25 µM Ad.1 and Ad.2, as previously published. Total PCR cycle was determined according to the amplification graph after the first 12 cycles. To remove primer dimers and larger fragment, double-sided bead purification was carried out using AMPure XP. Omni-ATAC-Seq
GEO Accession
GSM5642622