GSM9352529: glp-1, Naïve, timepointNC, replicate3; Caenorhabditis elegans; ATAC-seq
Frozen worm pellets were allowed to thaw on ice, then homogenized to release nuclei. Repeated centrifugation was performed to remove cellular debris from nuclei samples. Purified nuclei were immediately subjected to the ATAC-seq procedure. Nuclei were gently resuspended in 47.5 µl Omni-ATAC buffer (composed of 2X Illumina Tagment DNA (TD) buffer, 16.5 µl 1X PBS, 0.1% TWEEN20, and 0.01% Digitonin) and mixed with 2.5µl of Illumina Tagment DNA Enzyme (TDE1) (Cat#20034197). The mixture was incubated at 37°C on a thermomixer set to 500 rpm for 0.5-1 hour. Tagmented DNA was then purified using a MinElute PCR Purification Kit (Qiagen #28004) and eluted in 10µl. The purified DNA fragments were stored at -20°C until PCR amplification Fragmented DNA was purified using a MinElute PCR Purification Kit (Qiagen #28004) and PCR amplified using NEBNext Ultra II Q5 Master Mix (M0544S) for 12 cycles. Each sample utilized a common Adapter 1 primer (Ad1) and a unique index Adapter 2 (Ad2*) primer. Libraries were size-selected to retain DNA fragments between 100bp-600bp using Blue Pippin service, quantified using a Qubit, and quality-checked using a Bioanalyzer.