Methods
Bisulfite sequencing applies routine sequencing methods on bisulfite-treated genomic DNA to determine methylation status at CpG dinucleotides. Other non-sequencing strategies are also employed to interrogate the methylation at specific loci or at a genome-wide level. All strategies assume that bisulfite-induced conversion of unmethylated cytosines to uracil is complete, and this serves as the basis of all subsequent techniques. Ideally, the method used would determine the methylation status separately for each allele. Alternative methods to bisulfite sequencing include Combined Bisulfite Restriction Analysis and methylated DNA immunoprecipitation (MeDIP).
Methodologies to analyze bisulfite-treated DNA are continuously being developed. To summarize these rapidly evolving methologies, numerous review articles have been written.
The methodologies can be generally divided into strategies based on methylation-specific PCR (MSP) (Figure 4), and strategies employing polymerase chain reaction (PCR) performed under non-methylation-specific conditions (Figure 3). Microarray-based methods use PCR based on non-methylation-specific conditions also.
Read more about this topic: Bisulfite Sequencing
Famous quotes containing the word methods:
“Cold and hunger seem more friendly to my nature than those methods which men have adopted and advise to ward them off.”
—Henry David Thoreau (1817–1862)
“With a generous endowment of motherhood provided by legislation, with all laws against voluntary motherhood and education in its methods repealed, with the feminist ideal of education accepted in home and school, and with all special barriers removed in every field of human activity, there is no reason why woman should not become almost a human thing. It will be time enough then to consider whether she has a soul.”
—Crystal Eastman (1881–1928)
“We are lonesome animals. We spend all our life trying to be less lonesome. One of our ancient methods is to tell a story begging the listener to say—and to feel—”Yes, that’s the way it is, or at least that’s the way I feel it. You’re not as alone as you thought.””
—John Steinbeck (1902–1968)