title: GSM9075200: MDA-MB-231_ShACO2-NLS-MTS_R1; Homo sapiens; ATAC-seq
library_construction_protocol: Approximately, 1 X 106 cells were lysed with QIAzol lysis reagent, followed by the addition of chloroform. Aqueous and organic phases were later separated by centrifugation. Aqueous phases were transferred to a new tube, followed by the addition of 100% ethanol to it, and mixed by pipetting. The sample was later transferred into an RNeasy Mini spin column for collecting total RNA. On-column DNAse digestion was performed to remove residual genomic DNA. Nuclei were isolated from 50,000 cells. The samples were tagmented by resuspending nuclei in 2.5μL TDE1 transposase (Illumina; FC-121-1030), 25μL TD 2x buffer (Illumina; FC-121-1030), 22.5μL nuclease-free water, followed by incubation at 37°C for 30 minutes. The tagmented DNA fragments were purified by using a Qiagen MinElute PCR Purification kit. The purified DNA fragments were amplified by PCR (10 cycles) using KAPA Hi Fi 2X ready-mix (Roche). The amplified products were purified with AmpureXP beads (Beckman Coulter), followed by verification of the final library profiles by HSD1000 screentape on a Tapestation 4200 (Agilent). The libraries were quantitated using the KAPA Biosystems qPCR kit (Roche) and were pooled together in an equimolar fashion, following experimental design criteria. The resulting pool was then loaded into the appropriate NovaSeq Reagent cartridge and sequenced on a NovaSeq6000 following the manufacturer's recommended protocol (Illumina Inc.).