GSM4161105: Snf2h_WT_GO1_ATAC_rep2; Mus musculus; ATAC-seq
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).
GEO Accession
GSM4161105