ChIP-Atlas: SRX7141771

Antigen

Cell type Class: Gonad
Cell type: Oocytes
MeSH Description: Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM).

library_strategy: ATAC-seq
library_source: GENOMIC
library_selection: other
library_construction_protocol: As all libraries generated in this study involved low-input techniques, samples collected were directly put into lysis buffer for further library construction. Briefly, 100 fully grown oocytes nuclei or growing oocytes were collected. After washing with 0.2%BSA/PBS, oocytes or nuclei were added into 8 µL Tagmentation Mix (33 mM Tris-Acetate, 66 mM K-Acetate, 10 mM Mg-Acetate, 16% DMF, and 0.02% Digitonin) containing 0.5 µL of Assembled Tn5/Adapter (760 µg/mL unassembled Tn5 enzyme with 0.143 vol of 100 µM Tn5MEDS-A/B oligonucleotides). After tagmentation at 37 °C for 30 min, the reaction was stopped with freshly made Stop buffer (100 mM Tris pH8.0, 100 mM NaCl, 0.4% SDS, 40 µg/mL Proteinase K) and incubated at 55 °C for 16 hours. After quenching SDS by adding 5 µL 25% Tween-20, PCR was performed to amplify the library for 16 cycles using the following PCR conditions: 72 °C for 5 minutes; 98 °C for 5 minutes; thermocycling at 98 °C for 20 seconds, 63 °C for 30 seconds, 72 °C for 60 seconds; followed by 72 °C 5 minutes. After the PCR reaction, libraries were purified with the 1.6× SPRI beads (Beckman).