Determination of DNA Methylation of Imprinted Genes in Arabidopsis Endosperm

The overall goal of this procedure is to determine the DNA methylation status of imprinted genes in endosperm. This is accomplished by first emasculating, the female parent. The female parent is then pollinated with pollen of the male parent.

Next, the endosperm is dissected out of the cross pollinated seed. The final step of the procedure is to perform bi sulfate sequencing of endosperm DNA. Ultimately, results can be obtained that show the DNA methylation status of the maternal or paternal allele of an imprinted gene through DNA sequencing polymorphism in different ecotypes and bi sulfate sequencing.