title: CUT&Tag-FOS of human-chimp iPSC-derived tetraploid patterned induced neurons harvested at 2 hr post-KCl stimulation
design: 20µl of DNA was PCR amplified with 5X KAPA Hifi Buffer (Roche, cat #KK2101), 0.3mM dNTP (Roche, cat #KK2101), 1µl KAPA HiFi Polymerase (Roche, cat #KK2101), and primers with Illumina adapters in a 50µl reaction at 72°C for 5min, 98°C for 30s, 14 cycles of 98°C for 10s, 63°C for 30s, 72°C for 1min, and 72°C for 5min. DNA was purified using double-sided size selection (0.5X, then 1.2X) with Ampure XP beads (Thermo Fisher, cat #NC0110018). Briefly, 25µl of SPRI beads were added to the PCR reaction, incubated at room temperature for 8 min, and allowed to adhere to the magnet for 8 min. The supernatant was moved to a new tube and 65µl of SPRI beads were added. After 8 min incubation at room temperature, beads were allowed to adhere to the magnet for 8 min, the supernatant was discarded, and beads were washed twice with 200µl 80% EtOH for 30s each. Beads were resuspended in 53µl 10mM Tris-HCl, pH 7.5 and placed on the magnet, and the supernatant was moved to a new tube. The supernatant was then further cleaned by adding 55µl beads, incubated at room temperature for 8 min, washed twice with 200µl 80% EtOH for 30s each, and then eluted in 20µl 10mM Tris-HCl, pH 7.5. Libraries were quantified with the KAPA Library Quantification Kit (Roche, cat #07960140001).