ATAC-seq of Subclone 8 of Bridge Population 2a
Cells were trypsinized and washed twice with PBS. Then, 10,000 cells in 5L PBS were transposed in 42.5Lof transposition buffer (33mM Tris acetate buffer, 66 mM potassium acetate, 10mM magnesium acetate, 0.1% NP-40, 16% DMF, 0.004x protease inhibitor cocktail, ddH20 to 42.5uL) and 2.5L of TDE1 Illumina Tn5 transposase. The transposition reaction was conducted for 30 minutes at 37, followed immediately by DNA purification using the ZYMO DNA Clean and Concentrator 5 kit (Zymo Research). Cycle-determining qPCR was conducted to amplify libraries and stop amplification prior to saturation. The amplified libraries were purified using a ZYMO DNA Clean and Concentrator 5 kit and quantified by using Qubit 2.0 Fluorometer. The libraries were normalized and pooled based on qPCR analysis and were subsequently sequenced on an NovaSeq S1 instrument (Illumina) with 2 x 50 bp paired end configuration or a NextSeq instrument (Illumina) with 2 x 38 bp configuration at the Bauer Core Facility of Harvard University.