GSM9789411: ATAC-seq T2 H6 rep3; Vigna radiata; ATAC-seq
Nuclei were isolated from frozen leaf tissues. Tissue was ground in liquid nitrogen and resuspended in ice‑cold nuclei isolation buffer (25 mM Tris‑HCl pH 7.6, 10 mM MgCl2, 0.44 M sucrose, 0.1% Triton X‑100, 10 mM β‑mercaptoethanol, 2 mM spermidine, 0.1 mM PMSF, 1× protease inhibitor cocktail). The homogenate was filtered through a 40 μm cell strainer. Nuclear integrity was confirmed by DAPI staining. Approximately 50,000 intact nuclei were used for library preparation. Transposition was performed using Tn5 in a 50 μL reaction system at 37°C for 30 min. DNA fragments were purified using the QIAquick PCR Purification Kit (Qiagen). Libraries were amplified for 11–13 cycles using the TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme, China). Amplified products were purified with VAHTS DNA Clean Beads (Vazyme). Sequencing was performed on the Illumina NovaSeq 6000 platform to generate 150 bp paired‑end reads.