DNA Methylation Clinical Trial
Official title:
Comparison of the Methylation Pattern of DNA in Buccal Swabs From Different Time Points
A buccal swab will be taken using a cotton stick by rubbing it against the buccal mucosa.
Samples will be taken from the same participant at different timepoints.
DNA will be extracted from the buccal cells on the swabs using a commercial extraction kit
and will be quantified by a nano-drop spectrophotometer.
We will determine global and gene-specific methylation and hydroxymethylation alterations by
UPLC/MS/MS. Hydrolyzed DNA into individual nucleosides will be analyzed for the quantitative
measurement of 5-methylCytosine and 5-hydroxymethylCytosine on Triple Quadrupole UPLC/MS/MS
platform. Secondly, specific methylation in the CpG islands of tumor suppressor and promoter
genes, and genes involved in the oxidative stress pathways will be assessed by
PCR-pyrosequencing.
n/a
Observational Model: Cohort, Time Perspective: Prospective
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