Parental genomes can segregate to distinct blastomeres during the zygotic division resulting in haploid or uniparental diploid and polyploid cells, according to a study published in GenomeBiology.
]. For isolation of single cells from blastocyst-stage embryos, zygotes showing a multipolar division were cultured for 7 or 8 days, until they reached the blastocyst stage. Removal of the zona was followed by a wash step with TCM-199 with 10% FBS, and if present, large cells located in the perivitelline space were separated from the blastocyst. Cells of the blastocyst were further dissociated by a short incubation in trypsine-EDTA at 38.5°C, two wash steps in Ca-free PBS with 0.
Whole-genome amplified products were normalized to 150 ng/μL or 50 ng/μL before downstream processing with the Infinium HD assay super protocol . Single-cell, multi-cell sibling, and bulk parental or grandparental DNA were genotyped on BovineHD BeadChips . Subsequently, discrete genotypes, B-allele frequency values, and LogR values were exported using Ilumina’s GenomeStudio and fed to a modified version of the siCHILD algorithm [], siCHILD-bovine.
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