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:
“I think it is a wise course for laborers to unite to defend their interests.... I think the employer who declines to deal with organized labor and to recognize it as a proper element in the settlement of wage controversies is behind the times.... Of course, when organized labor permits itself to sympathize with violent methods or undue duress, it is not entitled to our sympathy.”
—William Howard Taft (1857–1930)
“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)
“The reading public is intellectually adolescent at best, and it is obvious that what is called “significant literature” will only be sold to this public by exactly the same methods as are used to sell it toothpaste, cathartics and automobiles.”
—Raymond Chandler (1888–1959)