GSM4388151: MOLM13 ATAC_rep2; Homo sapiens; ATAC-seq
MOLM-13 cells were subjected to cell lysis in cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630). Following 30 seconds of lysis, cells were centrifuged at 500 ×g for 10 minutes, 4°C. Lysed cell pellets were then subjected to the transposition reaction at 37°C for 30 minutes using Tn5 Transposes (Illumina Cat #FC-121-1030), followed by purification of transposed DNA using the Qiagen MinElute Kit. Transposed DNA fragment were amplified using NEBNext High-Fidelity 2x PCR Master Mix (New England Labs Cat #M0541) and Customized Nextera PCR primers under the following conditions: (Step 1) 72°C for 5 minutes, (Step 2) 98°C for 30 seconds, (Step 3) 98°C for 30 seconds, (Step 4) 63°C for 30 seconds, (Step 5) 72°C for 1 minute, and (Step 6) 4°C hold, with amplified library purified using Qiagen PCR Cleanup Kit prior to Illumina Sequencing. For PCR steps 3-5, transposed library was amplified with number of cycles based on quantitative PCR in order to stop amplification prior to saturation, thus reducing GC and size bias in PCR.
GEO Accession
GSM4388151