DNA Hydroxymethylation Analysis
DNA Hydroxymethylation - 5-hydroxymethylcytosine or 5'hmC - has gained attention in recent years as a potential important epigenetic factor in several areas, including the development of cancer, neurological disorders and stem cells. Although 5'hmC has been investigated intensively, its exact role is still not clear.
In the past, epigenetic studies have generally been focused on any kind of modification of the cytosine base. Cost effective technologies could not differentiate 5'mC and 5'hmC in one experiment. Now we can!
We already perform standard DNA Methylation services, and epigenetic analysis on samples derived from FFPE tissues. As an expansion of our DNA methylation profiling portfolio, we are also able to distinguish 5'hmC from 5'mC, using the Cambridge Epigenetix TrueMethyl™ Kit in combination with the Illumina Infinium HumanMethylation450K assay. Contact us for more information on this new protocol.
Our Hydroxymethylation Detection service includes:
- Unbiased Project Consultancy
- Sample Quality Control (QC)
- (ox)Bisulfite Conversion
- Bisulfite Conversion QC
- Illumina Infinium assay
- Data extraction and QC
Hydroxymethylation Analysis Assay
Hydroxymethylation detection uses the same methylation platform as our regular 5'mC profiling service. It is based on the Illumina infinium assay, using the human methylation450K (450K) beadchip. We distinguish 5'mC from 5'hmC by treating each sample with bisulfite and perform a proprietary oxidative step prior to the bisulfite conversionan to another aliquot of the same sample. After oxidation the 5'hmC is no longer bisulfite resistant and therefore converets into a T. This results in two data sets for differential methylation analysis. After substraction of the datasets you can easily see what part was hydroxymethylated.
DNA Methylation Analysis Service
The methylation service includes the following steps:Quality Control on the DNA samples, bisulfite treatment, quality control on the bisulfite converted samples, infinium assay, and primary data extraction with data quality control. This ISO 17025 accredited service (by GenomeScan) is performed following strict Standard Operating Procedures for optimal results.
For more detailed information on the service workflow, Infinium assay, sample requirements and more, download our DNA Methylation Guidelines.