GSM2752610: pre-MBT ATAC-seq rep1; Danio rerio; ATAC-seq
Staged embryos were lysed on ice for 10min (10mM Tris-HCl [pH 7.4], 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL CA-630). During the lysis, a 20 Gauge syringe was used to break the chorions and cells, followed by table top spin for 10 second. The supernatant was then centrifuged at 1300g 5 min 4℃ and nuclei was resuspend in 2X TD buffer, and incubated with Tn5 enzyme for 30 min at 55℃ (Nextera DNA Library Preparation Kit, FC-121-1031). The reaction was terminated by adding SDS to a final concentration 1%. Tn5 fragmented DNA was purified by Qiagen minElute column and PCR amplified by NEBNext High-Fidelity 2X PCR Master Mix (NEB, M0541L). The purified DNA have adaptes ligated and are ready for sequencing.
GEO Accession
GSM2752610