title: GSM8741254: COLO320-HSR cells YY1-CUT&Tag; Homo sapiens; OTHER
library_construction_protocol: Using the Hieff NGS® Multi-Cut &Tag Library Prep Kit for Illumina® (YEASEN), approximately 200,000 cells were collected, counted, and resuspended in 1× Wash Buffer, followed by the addition of ConA Beads. The mixture was incubated at room temperature for 10 minutes. Next, the cell-magnetic bead complex was resuspended in pre-cooled Primary Antibody Buffer containing lig3 (1:50), YY1 (1:50), H3K4me3 (1:50), and H3K27ac (1:50), and incubated at room temperature for 2 hours. After the incubation, a 1× Dig-300 Buffer containing Anti-mouse Tn5 and Anti-rabbit Tn5 was added to facilitate DNA transposition, which was performed at room temperature for 1 hour. Subsequently, transposase was activated, and the reaction was incubated at 37 °C for 60 minutes. Following transposition, a termination solution containing Proteinase K and a DNA spike-in mix was added, and the reaction was incubated at 55 °C for 30 minutes to digest proteins. Genomic DNA was then purified using DNA Extract Beads. The library was amplified using HiFi Amplification Mix in a PCR program consisting of an initial 3-minute extension at 72 °C, followed by 3 minutes of denaturation at 95 °C. The amplification cycle included 10 seconds of denaturation at 98 °C, 15 seconds of annealing at 60 °C, and a 1-minute extension at 72 °C. After amplification, the library was purified using DNA Selection Beads, and any residual impurities were removed by magnetic bead recovery and ethanol washing. The beads were air-dried and resuspended in ddH₂O to produce a library suitable for sequencing on the Illumina platform. Library quality was assessed using the Agilent 2100 Bioanalyzer.
identifiers.namespace: GEO
identifiers.id: GSM8741254_r1