GSM4699543: SN_rep1; Arabidopsis thaliana; ATAC-seq
For SN and VN ATAC-seq, ~50,000 FACS-purified nuclei were immediately resuspended in 25 μl transposase reaction mix (12.5 μl 2x TD buffer, 2.0 μl Nextera Tn5 [Illumina Cat #FC-121-1030] and nuclease-free water). For microspore ATAC-seq, FACS-purified spores were centrifuged at 800 g for 5 minutes at 4°C and the spore pellet resuspended in 500 μl lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 1 % IGEPAL CA-630). The microspores were pelleted once more at 500 g for 10 minutes at 4°C and ~50,000 added to a 25 μl transposase reaction mix (12.5 μl 2x TD buffer, 2.0 μl Nextera Tn5 [Illumina Cat #FC-121-1030] and nuclease-free water). The Tn5 reaction mixes were incubated at 37°C for 1 hour and tagmented DNA fragments recovered with a MinElute PCR Purification Kit (Qiagen). Sequencing-ready libraries were generated as described previously (Buenrostro et al., 2015). The libraries were sequenced on an Illumina Hiseq 2500 using 50 bp paired-end reads. All ATAC-seq profiles were generated with at least two independent biological replicates.
GEO Accession
GSM4699543