A simple Dot Blot Assay for population scale screening of DNA methylation

BioRxiv : the Preprint Server for Biology
Nelia LuvianoChristoph Grunau

Abstract

The study of epigenetic changes in natural and experimental populations has increased the need to find a cost-effective and high throughput method to analyze multiple samples to effectuate a population-wide screening to study epigenetic changes triggered by biotic or abiotic stress. One of the most studied epigenetic marks is global DNA methylation, its measurement is used as a first step to differentiate methylation between individuals. There is a wide range of methods designed to detect genome-wide 5 methyl-cytosine (5mC) that differ in sensitivity, price, level of expertise required, but as a general rule, require large amounts of DNA and are relatively expensive. This is a limit for the analysis of 5mC in a large number of individuals as a prerequisite to population-wide testing of methylation markers. In this work, we evaluated a method based on antibody recognition of 5mC to measure the DNA methylation level of individuals of the species Biomphalaria glabrata , the intermediate host of schistosomiases, a neglected tropical disease. We validated the method to complete a large screening in the genome of B. glabrata snails treated with a chemical inhibitor of DNA methylation; however, the method can be applied to any species...Continue Reading

Related Concepts

Australorbis glabratus
Biological Markers
Cytosine
DNA
Enzyme Inhibitors
Enzyme-Linked Immunosorbent Assay
Genome
Methylation
Schistosomiasis
Snails

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