ChIP-Atlas: SRX4065618

Antigen

Cell type Class: Embryo
Cell type: Morula
MeSH Description: An early embryo that is a compact mass of about 16 BLASTOMERES. It resembles a cluster of mulberries with two types of cells, outer cells and inner cells. Morula is the stage before BLASTULA in non-mammalian animals or a BLASTOCYST in mammals.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: SCNT embryos were generated from Sertoli cells of Xist heterozygous knockout pups in BDF1 x PWK hybrid genetic background through Kdm4d mRNA injection. IVF embryos were generated using BDF1 oocytes and PWK sperm. Fifty to sixty IVF or SCNT embryos at the morula stage were collected at 72 hpa and pooled as a single sample with biological replicates. After digestion of zona pellucida by acid Tyrode's solution, polar bodies were removed by brief treatment with 0.25% trypsin 1 mM EDTA followed by several times of quick pipetting. The embryos were then washed three times with PBS containing 0.2% BSA (Merck # 12657) and processed for chromatin isolation and fragmentation by Micrococcal Nuclease (MNase: New England Biolabs # M0247S). 10 % of MNase-treated chromatin solution was used as input. Rabbit anti-H3K27me3 antibody (Diagenode # C15410069) was used for immunoprecipitation. Sequencing libraries were constructed using NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs # E7645S) following manufacturer's instructions.