GSM3504934: e11.5_mouse_distal_forelimb_HOX13KO_ATACseq_rep2; Mus musculus; ATAC-seq
Dissection of proximal and distal of forelimb buds from wild type embryos and distal forelimb buds of Hox13-/- embryos 7 were performed at E11.5. All samples for ATAC-seq were processed as previously described 15. Briefly, 50 000 cells were washed in PBS and incubated on ice for 30 minute in a hypotonic cell lysis buffer (0.1% w/v sodium citrate tribasic dehydrate and 0.1% v/v Triton X100) centrifugated (5 minutes at 2000g at 4°C). Cells were then incubated 30 minutes on ice in cell lysis buffer (10mM Tris-HCl, pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% v/v IGEPAL CA-630. After centrifugation (5 minutes at 2000g at 4°C) the resulting pellet of nuclei was re-suspended in transposase Master Mix (1.25 µl 10x TD buffer, 5 µl H2O and 6.5 µl of Tn5: Illumina Nextera Kit; FC-121-1031) and incubated for 30 minutes at 37°C. Samples were purified using MinElute PCR purification column (Qiagen). The eluted DNA was enriched and barcoded for multiplexing of samples using Nextera barcodes by PCR using Phusion kit. Libraries were prepared according to Illumina's instructions. Libraries were sequenced on the Illumina Hi-seq 2500 following the manufacturer's protocols to obtain 50bp paired end reads.
GEO Accession
GSM3504934