Leiden DNA Methylation Analysis: Updated 12/2012

Background: The following procedure describes the detailed DNA methylation analysis process of liquid DNA samples by methylation-sensitive Southern blot performed at the Leiden University Medical Center. Detailed DNA methylation studies showed D4Z4 hypomethylation on chromosomes 4q and 10q at the FseI restriction site in patients with FSHD2.
Prior to the FseI digestion DNA samples need to be purified to guarantee a complete digestion using this methylation sensitive restriction enzyme (see protocol). This analysis simultaneously measures the D4Z4 methylation level on both chromosomes 4q and 10q. The methylation value at D4Z4 can be obtained by comparing the ratios of signal intensities between bands representing the unmethylated and methylated D4Z4.

Relevant References:

1. Lemmers RJ, Tawil R, Petek LM, et al. Digenic inheritance of an SMCHD1 mutation and an FSHD-permissive D4Z4 allele causes facioscapulohumeral muscular dystrophy type 2. Nat Genet. 2012 Nov 11;44(12):1370-4.


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