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:
“The methods by which a trade union can alone act, are necessarily destructive; its organization is necessarily tyrannical.”
—Henry George (1839–1897)
“We can best help you to prevent war not by repeating your words and following your methods but by finding new words and creating new methods.”
—Virginia Woolf (1882–1941)
“I believe in women; and in their right to their own best possibilities in every department of life. I believe that the methods of dress practiced among women are a marked hindrance to the realization of these possibilities, and should be scorned or persuaded out of society.”
—Elizabeth Stuart Phelps (1844–1911)