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EpiTect Methyl II PCR Arrays

EpiTect Methyl II PCR Arrays allow the simultaneous DNA methylation profiling of a panel of 22 or 94 genes promoters using restriction enzyme based MethylScreen™ technology. Genes are carefully selected based on their reported methylation statusin a variety of experimental settings. These arrays allow correlation of CpG island methylation status with biological phenotypes or disease outcomes. Both EpiTect Methyl II Signature PCR Arrays (22 genes) and EpiTect Methyl II Complete PCR Arrays (94 genes) are available for human, mouse or rat studies in 96- or 384-well formats.

With a simple restriction enzyme digestion followed by real-time PCR, you can simultaneously analyze the DNA methylation levels of 22 or 94 different disease- or pathway-focused genes.

Why EpiTect Methyl II PCR Arrays?

  • Complete System
    Sample Prep to Data Analysis using QIAGEN DNA Isolation Kits and Excel-based data analysis templates
  • High Throughput without Bisulfite Conversion-
    Analyze 22 or 94 genes per sample without bisulfite conversion
  • Guaranteed Performance
    Wet Bench tested qPCR Assays and Optimized Restriction Enzyme Digest Conditions

Disease- and Pathway-Focused DNA Methylation PCR Arrays:

Disease-focused Pathway-focused
Breast Cancer Apoptosis  
Cancer miRNA Cell Cycle
Colon Cancer Cytokine Production
Epithelial to Mesenchymal Transition (EMT)   DNA Repair 
Gastric Cancer Homeobox (HOX) Genes
Leukemia & Lymphoma   Inflammatory Response and Autoimmunity
Liver Cancer Mental Disorders
Lung Cancer Notch Signaling Pathway
Melanoma Polycomb & Trithorax Complexes
Prostate Cancer Stem Cell Transcription Factors
Tumor Suppressor Genes Stress & Toxicity
T Cell and B Cell Activation
T Helper Cell Differentiation
Tumor Suppressor Genes
Toll Like Receptor Signaling
TGF-β/BMP Signaling  
Wnt Signaling  

Principle

The method is based on the detection of remaining input DNA after cleavage with a methylation-sensitive (MSRE) and/or a methylation-dependent (MDRE) restriction enzyme. These enzymes will digest unmethylated and methylated DNA, respectively. Following digestion, the remaining DNA is quantified by real-time PCR in each individual enzyme reaction using primers that flank a promoter (gene) region of interest. The relative fractions of methylated and unmethylated DNA are subsequently determined by comparing the amount in each digest with that of a mock (no enzymes added) digest using the ΔCt method. The reliability and simplicity of the procedure make this technology highly suited for semi-high-throughput DNA methylation profiling and biomarker discovery for various research fields, such as stem cell differentiation and development.

How It Works

Input genomic DNA is aliquoted into four equal portions and subjected to mock (Mo), methylation sensitive (Ms), methylation dependent (Md), and double (Msd) restriction endonuclease digestion. After digestion, the enzyme reactions are mixed directly with qPCR master mix and aliquoted into a PCR Array plate containing pre-dispensed primer mixes. Run real time PCR using specified cycling conditions. Finally, copy and paste raw Ct values to data analysis excel sheet which automatically calculates the relative amount of methylated and unmethylated DNA fractions. Learn More.

Resources:

EpiTect Methyl II PCR Array Manual

EpiTect Methyl II DNA Restriction Kit
To prepare DNA samples for EpiTect Methyl II PCR Arrays or qPCR Primer assays of choice

RT² SYBR® Green qPCR Master Mixes
Instrument-specific PCR master mixes insure high-efficiency, sequence-specific amplification

Data Analysis Software (Excel template)

Attend a Webinar to have dialogues with our Application Scientists

The EpiTect Methyl II PCR Arrays use the MethylScreen™ Technology provided under license from Orion Genomics, LLC.

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