GSM8605837: ExC_Foxi1MO-st10; Xenopus laevis; ATAC-seq
Caps were transferred from MBS plates into a 1.5 mL low-bind microcentrifuge tube (Eppendorf, cat. No. 0030108051) containing 1 mL of ice-cold 1X PBS. Samples were spun at 500 g at 4 ºC in the centrifuge for five minutes before removing the PBS and repeating the wash step with fresh ice-cold 1X PBS. 50 µL of ice-cold lysis buffer (10 mM Tris pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% (w/v) Igepal CA-630) and pipetted to break up samples. Samples were centrifuged at 500 g for 10 min at 4 ºC and pellets were resuspended in 25 µL TD Buffer, 2.5 µL TDE1 Enzyme and 22.5 µL Nuclease-Free water (Illumina, cat. No 20034198). Samples were pelleted to mix and incubated on a ThermoMixer at 37 ºC, 700 rpm for 30 min. Following incubation, the samples were cleaned with MinElute Reaction Cleanup Kit (Qiagen, cat. No. 28204), following manufacturer instructions and eluted into 11 µL Buffer EB. Libraries were prepared with the ATAC-seq Kit (Active Motif, cat. No. 53150)