title: GSM4388150: MOLM13 ATAC_rep1; Homo sapiens; ATAC-seq
library_construction_protocol: 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.
attributes.tag: GEO Accession
attributes.value: GSM4388150