Methylation Analysis by Sequencing
DNA methylation is involved in the regulation of many cellular processes, including X chromosome inactivation, chromosome stability, chromatin structure, embryonic development, and transcription. Bisulfite sequencing is the gold standard for validating methylation experiments, as only sequencing provides direct detection of methylation events as well as information across the entire region. One of the key steps in this process, bisulfite DNA conversion, allows precise analysis of methylation in the target region by converting all non-methylated cytosines into uracils (methylated cytosines remain unchanged).
Two sequencing workflows can be use to determine the methylation status:
Step-by-Step Guide to Methylation Analysis
DNA extraction is a critical first step in the experimental workflow of DNA sequencing-based methylation analysis. The overall quality, accuracy and length of the DNA sequence read can be significantly affected by characteristics of the sample itself, and the method chosen for nucleic acid extraction. Ideal methods will vary depending on the source or tissue type, how it was obtained from its source, and how the sample was handled or stored prior to extraction.
Recommended Products: DNA Isolation
Promoter Hypermethylation in Thyroid Cancer Samples Publication:
Innovations Newsletter - Issue 09, Oct 2008