1-cell (5 h post insemination, between pronucleus (PN) stage II and early stage III)
genotype
SRPK1 knock-down
Sequenced DNA Library
library_strategy
ATAC-seq
library_source
GENOMIC
library_selection
other
library_construction_protocol
After removing the zona pellucida, fertilized eggs were washed with PBS. ATAC-seq libraries were generated using omni-ATAC-seq developed by Corces et al. with some modifications. Briefly, fertilized zygotes were incubated in cold resuspension buffer (10 mM Tris-Cl pH 7.4, 10 mM NaCl, and 3 mM MgCl2) containing 0.1% NP-40, 0.1% Tween-20, and 0.01% digitonin on ice for 3 min and then washed with cold resuspension buffer containing 0.1% Tween-20. Tagmentation was next performed with Tn5 transposon enzyme (Illumina) in tagmentation buffer (20 mM Tris-Cl pH 7.4, 10 mM MgCl2, 20% dimethyl foramide, 0.1% Tween-20, and 0.01% digitonin) for 30 min at 37°C in a thermomixer with shaking at 1,000 rpm. Tagmentated DNA was purified using DNA Clean & Concentrator-5 Kit (Zymo Research). PCR was performed to amplify each library for 14 cycles.