flank ATAC-seq, biological replicate 2
Tegu lizard embryos (DH29-30, mouse E11.5 equivalent) were dissected in PBS to obtain samples from fore- and hindlimbs, fore- and midbrain, heart, liver, lateral flank tissue (between fore and hindlimb buds), and the remaining embryonic material (here refered to as 'embryo'). Tissue samples were pooled from at least 10 embryos, and frozen in liquid nitrogen after dissection. ATAC was performed on each tissue in two biological replicates, following protocol from Buenrostro et al. 2013. In short, we ground approximately 4mg of tissue/sample on lysis buffer, spun for 10min at 400g, discarded supernatant, resuspended nuclei in ATAC mix (Illumina Nextera), and incubated for 30min at 37oC. Library amplification was done for 10 PCR cycles, using Illumina Nextera reagents. Libraries were quantified with Fragment Analyzer (Advanced Analytical) and KAPA (KAPA Biosystems) and equimolar pooled for sequencing. All libraries were sequenced on Illumina HiSeq 2500, to obtain 2x75 bp reads.