EpiTect Methyl II PCR Array Service
For pathway or disease-focused profiling of regional DNA methylation using MethylScreen technology
The EpiTect Methyl II PCR Array Service offers DNA methylation profiling services using the EpiTect Methyl II PCR Array system. This innovative technology enables fast and accurate detection of CpG island DNA methylation profiling of individual genes, as well as disease- or pathway-focused gene panels. The EpiTect Methyl II PCR Array Service uses MethylScreen technology provided under license from Orion Genomics, LLC.
The EpiTect Methyl II PCR Array Service is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
EpiTect Methyl II PCR Arrays rely on the differential cleavage of target sequences by 2 different restriction endonucleases whose activities require either the presence or absence of methylated cytosines in their respective recognition sequences. As the relative amount of DNA remaining after each enzyme digestion is measured by real-time PCR, the methylation status of individual genes and the methylation profile across a gene panel are reliably and easily calculated. The high yield of DNA from the restriction digests and PCR amplification allow the analysis of smaller, more heterogeneous samples.
The genes included in EpiTect Methyl II PCR Arrays have been carefully selected based on their reported hypermethylation in a variety of experiments. Profiling your biological DNA samples with these arrays will allow you to correlate CpG island methylation status with biological phenotypes or disease outcomes, and the results can provide insights into the molecular mechanisms and biological pathways critical for disease development.
EpiTect Methyl II PCR Arrays are available in both 96- and 384-well plate formats enabling simultaneous analysis of 24 or 96 pathway- or disease-specific gene panels. The signature panels contain 24 genes and are arranged in either 96- or 384-well plates for characterizing all 4 restriction digests from either 1 or 4 different DNA samples, respectively. The complete panels contain 96 genes and are arranged in 96- or 384-well plates for characterizing all 4 restriction digests from one DNA sample either on 4 plates or all on the same plate, respectively.
Four different restriction digests are carried out on a DNA sample: methylation-sensitive enzyme, methylation-dependent enzyme, double digest with both enzymes, and a mock digest with no enzyme. Next, quantitative real-time PCR of genes of interest detect the amount of DNA remaining in each reaction. The methylation status — methylated, hypermethylated, or unmethylated — is calculated from these amounts.
Choose from the following predesigned cancer or pathway-focused arrays (see Table), or select EpiTect Methyl II Custom PCR Arrays and define your own gene list. Proprietary algorithms reliably predict promoter CpG islands and select the best target promoter region to analyze. Rigorous qPCR primer verification and quality control manufacturing process guarantee reliable assay performance.
For instructions regarding sample submission and shipping, please contact QIAGEN Technical Services. Samples accepted are cells or tissue or total DNA. For cells and tissue, a DNA isolation service is available.
For all samples
Please note the minimum amount of starting materials required (see below for requirements for each sample type). If you have less than the minimum amount or your previously isolated samples have been stored differently than we recommend, please contact QIAGEN Technical Services before submitting your samples.
Provide at least 0.5 million cells for each sample, and specify the number of cells on the submission form. Snap freeze the cell pellets in liquid nitrogen, store samples frozen at –80°C, and ship on dry ice.
Provide at least 50 mg of tissue for each sample, and specify the weight of the tissue on the submission form. Snap freeze tissues in liquid nitrogen, store samples frozen at –80°C, and ship on dry ice.
Purified genomic DNA
Purified genomic DNA is required for methylation analysis. A genomic DNA isolation service is currently also available, please inquire.
Genomic DNA isolated using a standard genomic DNA isolation kit, such as the DNeasy Blood & Tissue Kit, which is compatible with methylation profiling. We recommend 2 µg DNA per sample (in DNase-free water or TE buffer) at a concentration ≥30 ng/µl. A260/A280 ratio of ≥1.7 and A260/A230 ratio of ≥1.7 is required. Customers should check sample integrity by agarose gel electrophoresis before sending samples to QIAGEN. Sample gel pictures, if available, should accompany the submitted samples.
Samples should also be accompanied by a sample submission sheet. Excel format is preferred for sample submission, and the spreadsheet should contain the following information: sample ID, concentration, A260/A280, A260/A230, total volume, total DNA amount.
Data in an electronic form can be downloaded from a secure FTP website or delivered via email. A complete data and experiment report includes:
The EpiTect Methyl II PCR Array Service is highly suited for methylation profiling of pathway or disease-focused gene panels and discovery and development of biomarkers important to stem cell growth and differentiation, cancer, and other human diseases.