GSM8464301: ATAC_ddmsn_A; Saccharomyces cerevisiae; ATAC-seq
The main steps of our ATAC-seq protocol are outlined here; for detailed information, please refer to the Methods section. Cells were cultured as described in the MPBA protocol until reaching an OD600 of 4, then spheroplasted using lyticase. The resulting spheroplasts were treated with RNase-A. Subsequently, a tagmentation reaction was performed using in-house produced Tn5 transposase loaded with Tn5MEDS-A/B oligonucleotides. The reaction was terminated after 1 hour using SDS and Proteinase K The eluted samples were used as a template for a PCR reaction for amplifying Tn5 tagmented fragments and sample barcoding (2× Kappa HIFI [Roche], 5 μL nextera primers F: i5, R: i7; 12 cycles; see primer list in Supplementary Table 4). Next, samples were 0.65x reversed SPRI cleaned and resuspended in 22 μL elution buffer (10mM Tris-HCl, pH 8), then concentrations and library quality were validated using Qubit and tape station (Agilent). ATAC-seq; Libraries were sequenced using the NovaSeq 6000 machine. Runs were performed with the S1 kit (20028319), parameters: R1 - 61 cycles, Index1 - 8 cycles, Index2 - 8 cycles, R2 - 61 cycles.