EpiTect Control DNA and Control DNA Set
For control experiments during methylation analysis
- Convenient, ready-to-use, quality-controlled DNA
- Bisulfite-converted DNA for control experiments
- Suitable for all methylation analyses
Cat. No. / ID: 59695
Accessing epigenetic information is of prime importance for many areas of biological and medical research — particularly oncology, but also stem cell research and developmental biology. However, the analysis of changes in DNA methylation is challenging, due to the lack of standardized methods for providing reproducible data particularly from limited sample material. With its newly introduced EpiTect solutions, QIAGEN makes available standardized, pre-analytical and analytical solutions from DNA sample collection, stabilization and purification, to bisulfite conversion and real-time or end-point PCR methylation analysis or sequencing (see figure " Standardized workflows in epigenetics").
EpiTect Control DNAs are suitable for use in control reactions for all types of methylation analyses, including:
EpiTect Methylated Control DNA and EpiTect Unmethylated Control DNA (both pre-bisulfite converted) were mixed to give 100%, 90%, 50%, 10%, and 0% methylated DNA. EpiTect MethyLight Assays for the human PITX2 gene were run in triplicate, using 10 ng of each DNA sample. The results show that EpiTect Control DNA can be used as a methylation standard for the quantification of unknown DNA samples.
|Applications||End point Methylation Specific PCR (MSP), real-time methylation specific PCR|
|Number of reactions||for 1000 control PCRs|
|Concentration||10 ng/µl (1 µg in total) excepted the unmethylated control DNA: 50 ng/µl (10 µg in total)|
|Type of DNA||Primer for unmethylated & unconverted genomic DNA||Primer for unmethylated & bisulfite converted target DNA||Primer for methylated & bisulfite converted target DNA|
|Unmethylated & untreated control DNA||PCR product||NO PCR product||NO PCR product|
|Unmethylated & bisulfite converted control DNA||NO PCR product||PCR product||NO PCR product|
|Methylated & bisulfite converted control DNA||NO PCR product||NO PCR product||PCR product|
|No template control||NO PCR product||NO PCR product||NO PCR product|
No, the EpiTect PCR Control DNA Set is derived from human DNA. No primers for mouse or rat will bind.
The unmethylated and unconverted human control DNA of the EpiTect PCR Control DNA Set allows to check that primers designed for the specific detection of unmethylated and converted DNA (U-converted DNA), and for methylated, converted DNA (M-converted DNA) do not bind to untreated genomic DNA.*
In case bisulfite conversion was not complete, leaving certain unmethylated C residues unconverted, false positives would result if the primer specific for M-converted DNA binds to untreated gDNA.
This control DNA can also be used to check conversion efficiency during bisulfite treatment.
*(Summary of principle: Methylation of DNA occurs on cytosine residues, especially on CpG dinucleotides enriched in small regions of DNA. Incubation of target DNA with sodium bisulfite, using e.g. EpiTect Bisulfite Kits, results in conversion of unmethylated cytosine residues into uracil, leaving methylated cytosines unchanged).