GSM7999943: KAS-seq_GVBD_biol rep2; Mus musculus; OTHER
Oocytes were labeled with N3-Kethxal (5 mM), and labeled genomic DNA (gDNA 9.5 µL) was subsequently extracted using the Quick-DNA Microprep Plus Kit (Zymo D4074) After labeling, gDNA was fragmented using Tn5 transposase (Vazyme TD501).Following fragmentation, biotinylation labeling was performed using DBCO-PEG4-biotin (20 mM, DMSO solution, Sigma 760749). The biotinylated DNA was then cleaned up using DNA Clean & Concentrator-5 kit (Zymo, D4013), and 5 µL of labeled DNA was retained as input, while the remaining 50 µL of DNA was used for enrichment.The Dynabeads™ MyOne™ Streptavidin C1 (Thermo 65001) were mixed with the 50 µL fragmented DNA obtained from the previous steps and rotated slowly at room temperature for 15 min.DNA-conjugated beads and their corresponding inputs were used for library PCR using i5 and i7 index primers (Vazyme TD205) and VAHTS HiFi Amplification Mix (Vazyme N616). The PCR reactions were initiated with a 5-min incubation at 72°C, followed by 10 min at 95°C, and then amplified for 17 cycles (10 s at 98°C, 30 s at 60°C, 1 min at 72°C). Finally, the libraries were cleaned up by using DNA Clean & Concentrator-5 kit (Zymo, D4013).