GSM5931494: xenopus tropicalis sperm-ATAC_rep2; Xenopus tropicalis; ATAC-seq
Male Xenopus tropicalis were injected with 150 U HCG. After 2 hours, two X. tropicalis testes collected in 1 ml 1xMBS and homogenized using razor blades. The solution was filtered with 40um strainer and its volume was brought down to 14ml. Then, through twice 180G horizontal centrifugation, we can obtain pure sperm in the supernatant. About 50000 sperms were lysed in lysis buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2 and 0.5% NP-40) for 10 min on ice to prepare the nuclei. Then, nuclei were spun at 1500g for 5 min to remove the supernatant. Nuclei were then incubated with the Tn5 transposome and tagmentation buffer at 37 °C for 30 min (Vazyme，TD501).  Following incubation, sperm were treated with Proteinase K at 63°C for 2 hr, and gDNA was purified with the 1.2× DNA clean beads. PCR was performed to amplify the library for 12 cycles and libraries were purified with the 1.2×DNA clean beads. Amplified ATAC libraries were sequenced on the Illumina HiSeq X10 platform.