Detection and Technology
Detecting Methylated Septin9
Cytosine residues in the v2 region of the SEPT9 gene may become methylated in colorectal cancer tissues. When DNA isolated from plasma samples is treated with a high concentration of bisulfite, unmethylated cytosines are converted to uracil while methylated cytosines remain unchanged (Figure 1). As a consequence of treatment, the DNA sequence is altered based on methylation status and can be analyzed by Real-Time PCR amplification (Figure 1).
Figure 1: Detecting DNA Methylation
HeavyMethyl® Core Technology
The Epigenomics’ HeavyMethyl core technology combines the use of primers that amplify the target biomarker regardless of methylation status, with a blocking oligonucleotide to suppress the amplification of unmethylated DNA, and a methylation-specific probe to detect the amplified methylated sequence (Figure 2). The proprietary HeavyMethyl core technology enables detection of low copy number tumor DNA in a background of non-tumor DNA in plasma.
FIGURE 2: HeavyMethyl® Real-Time PCR