Methylation Analysis
Methylation of Cytosine-Guanine (CpG) dinucleotides in genomic DNA has been correlated to gene silencing, and is regarded as one of the most significant epigenetic events. In particular, CpG methylation in the promoter region of certain genes has been observed as the earliest and most frequent alteration in some cancers by causing inactivation of tumor suppressors.
Analysis of Methylation using Bisulphite Sequencing
Bisulphite conversion of unmethylated Cytosine is an easy and widely accepted method for detecting methylated CpGs. Furthermore, sequencing bisulphite treated DNA is the gold standard technique for methylation detection. SeqWright offers bisulphite treatment, PCR, DNA sequencing, and comparative analysis as an integrated solutions package for detecting the presence of Cytosine methylation. Service includes:
- DNA extraction (including from paraffin-embedded tissues)
- Preliminary PCR and sequencing of genomic DNA
- Bisulfate conversion of unmethylated Cytosines
- Primers designed for amplifying only converted DNA for improved results
- Subcloning of PCR products or direct sequencing
- ABI Prism™ 3730xl DNA Sequencers
- Enhanced sequence delineation abilities through Cytosine deficient templates
- Detection by TaqMan® assay available through our qPCR Services
- Sequence comparison and analysis of 5MeCpGs
Genome-Wide Methylation Analysis on an Affy 100K/500K Chip
SeqWright offers genome-wide scans for detecting hyper/hypo-methylation on an Affymetrix 100K or 500K chip. Our proprietary technique takes advantage of certain methylation-sensitive enzymes combined with a novel screening and detection technology developed by our Microarray division. Targeted identification and validation of methylated CpGs is subsequently performed using DNA sequencing.
|
