title: GSM8465744: AP2XII 8 CUT&Tag 2; Toxoplasma gondii; OTHER
library_name: GSM8465744
library_construction_protocol: For CUT&Tag Library construction, the library was performed using the NovoNGS CUT&Tag 4.0 High-Sensitivity Kit for Illumina (Novoprotein, Suzhou, China) according to the manufacturer's instructions. Briefly, fresh tachyzoites were bound to activated concanavalin A beads (10 μL/sample) and incubated for 10 min at room temperature. The mixture was resuspended and incubated with primary antibody (1:50, mouse anti-HA) at 4°C overnight. After several washes, the parasites were incubated with secondary antibody (1:100, goat anti-mouse IgG) for 1 h at room temperature. The parasites were then resuspended with pAG-Transposome buffer and incubated for 1 h at room temperature on a rotator. Tagmentation was stopped by MgCl2 treatment and DNA extraction was performed using DNA extract beads (Novoprotein, Suzhou, China). For CUT&Tag Library construction, Illumina sequencing libraries were generated by PCR amplification using specific adaptors according to the manufacturer's recommendations (NovoNGS CUT&Tag 4.0 High-Sensitivity Kit for Illumina B box, Novoprotein, Suzhou, China).
identifiers.namespace: GEO
identifiers.id: GSM8465744_r1