title: GSM8553455: H3K4me3_mGFP_Control_2C_rep2; Mus musculus; OTHER
library_name: GSM8553455
library_construction_protocol: Low-input CUT&Tag: For more information, see the methods in the accompanying paper. Briefly, late 2-cell embryos with intact zona pellucida were collected 48h post-hCG. They were permeabilized using a Triton-X buffer and incubated overnight at 4 °C with a primary antibody against H3K4me3. After this, the embryos were incubated at room temperature for 30 minutes with a secondary antibody. Next, the pA-Tn5 adaptor complex was added, and the mixture was incubated for 1 hour at room temperature. Tagmentation was then performed for 1 hour at 37 °C in a MgCl₂ buffer. DNA was extracted by incubating the samples for 1 hour at 58 °C in a 0.1% SDS buffer. SDS was neutralized with Triton-X, and the PCR reaction was conducted directly on the sample. Following 18 cycles of PCR amplification with NEBNext High-Fidelity 2X PCR Master Mix, the PCR product was purified and size-selected using AMPure XP. See methods of the accompanying paper for more information.
identifiers.namespace: GEO
identifiers.id: GSM8553455_r1