GSM5006002: ATAC_seq_Meristems_rep2; Solanum lycopersicum; ATAC-seq
Six biological replicates of 1g of freshly collected anantha inflorescences (meristematic enriched tissue) were ground in ice-cold isolation buffer (300 mM sucrose, 20 mM Tris pH 8.0, 5 mM MgCl2, 5 mM KCl, 5 mM beta-mercaptoethanol, 35% glycerol, 0.2% Triton X-100). After grinding, the suspension was filtered through a series of cell strainers (the smallest being 70 mm). Samples were further disrupted using a Dounce homogenizer, and then were washed four times with ice-cold isolation buffer. The enriched nuclei samples were resuspended in Resuspension Buffer (50 mM Tris pH 8.0, 5 mM MgCl2, 5 mM beta-mercaptoethanol, 20% glycerol). For the transposase reaction 100,000 nuclei were resuspended  in 22.5 ml freezing buffer (5 mM MgCl2, 0.1 mM EDTA, 50 mM TRIS pH 8.0, 40% glycerol), and were mixed with  2X DMF Buffer (66 mM Tris-acetate (pH 7.8), 132 K-Acetate, 20 mM Mg-Acetate, 32% DMF) and 2.5ul Transposase enzyme from Illumina Nextera Kit (Catalog No. FC-121-1031), and incubated at 37°C for 30 min. Tagmented DNA was isolated using NEB Monarch PCR Purification kit (Catalog No. NEB #T1030).  Next-generation sequencing (NGS) libraries were amplified in two steps (Buenrostro et al., 2015) using KAPA HiFi HotStart ReadyMix PCR kit (Catalog No. KK2601). NGS libraries were PE50 sequenced on an Illumina MiSeq SY-410-1003 instrument.