EpiTect Control DNA and Control DNA Set

For control experiments during methylation analysis

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

EpiTect PCR Control DNA Set (100)

Cat. No. / ID: 59695

Human control DNA set (containing both bisulfite converted methylated and unmethylated DNA and unconverted unmethylated DNA) for 100 control PCRs

Inquire

EpiTect Control DNA and Control DNA Set are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Convenient, ready-to-use, quality-controlled DNA
Bisulfite-converted DNA for control experiments
Suitable for all methylation analyses

Product Details

EpiTect Control DNAs are ready-to-use, completely methylated or completely unmethylated bisulfite converted DNAs, and untreated, unmethylated genomic DNA, for standardized and reliable control reactions for methylation analysis. The methylated, bisulfite converted DNA as well as unmethylated, bisulfite converted and unconverted DNA are stored in EB Buffer (10 mM Tris Cl) in a ready-to-use solution.

Performance

EpiTect Control DNAs are highly suited for use as quantification standards for probe-based real-time PCR methylation analysis (see figure "") and high-resolution melting (HRM) methylation analysis (see figure "").

See figures

Real-time PCR standards.

HRM quantification standards.

Principle

The EpiTect Control DNA Set contains all control DNAs needed for standardized and reliable methylation control reactions in a ready-to-use kit format . In methylation-specific PCR, for example, control reactions must be carried out to ensure that PCR probes and primers bind specifically to methylated or unmethylated DNA (see figure ). These reactions require various concentrations of completely bisulfite converted control DNA that is fully methylated or fully unmethylated (see figures and ""). Additionally, mixtures of these control DNAs can serve as quantification standards when determining degree of methylation in HRM and probe-based methylation experiments.

See figures

Use of EpiTect Control DNA in PCR for methylation analysis.

EpiTect Control DNA Pyrogram.

EpiTYPER MALDI-TOF analysis of WBA DNA.

Procedure

Standardized workflows in epigenetics

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").

See figures

Standardized workflows in epigenetics.

Applications

EpiTect Control DNAs are suitable for use in control reactions for all types of methylation analyses, including:

Evaluating primer specificity for MSP
Quantification standard for HRM and MethyLight PCR
Evaluating primer and probe specificity for MethyLight PCR
Checking efficiency of bisulfite conversion reactions

Supporting data and figures

Real-time PCR standards.

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.

Specifications

Features	Specifications
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)

Resources

For performing 100 control reactions
for methylation analysis using Human
control DNA

FAQ

Expected PCR results when using the EpiTect Control DNA Set in combination with the EpiTect MSP Kit can be found in the table below:

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

FAQ ID -2011

Yes, the EpiTect Control DNA and Control DNA Set can be used as standards in real-time methylation specific quantification. For data, please see our EpiTect MethyLight PCR Product Profile.

FAQ ID -2006

No, the EpiTect PCR Control DNA Set is derived from human DNA. No primers for mouse or rat will bind.

FAQ ID -2008

We have data for DNA converted with EpiTect Bisulfite Kits showing that the bisulfite treated DNA is stable for 3 years at -20°C before use in methylation specific PCR with the EpiTect MSP Kit, or the EpiTect MethyLight PCR Kit.

FAQ ID -2002

The genomic DNA is derived from disease free human whole blood from multiple anonymous donors. The DNA is pooled after purification to yield a mixture of individuals (male/female) and the whole genome is represented. The whole genome is copied stripping the resultant DNA of methylation.

The methylated DNA control is methylated in vitro using SssI methylase. The methylation efficiency is high but not guaranteed to be 100% for every position. Although methylated under optimized conditions for the enzymatic reaction, some positions are as low as 85% as determined by Pyrosequencing. These sites of lower methylation are not random but rules to predict these low positions are undetermined.

QIAGEN checks the consistency of EpiTect Control DNA for methylation and bisulfite conversion using an very sensitive real time MSP-PCR assay. QIAGEN performs several realtime PCR tests to evaluate the bisulfite conversion efficiency as well as the methylation efficiency of the DNAs in the Control DNA Set. We use test systems specific for genomic DNA to detect unconverted DNA or for converted and unmethylated DNA to detect converted but unmethylated DNA, as well as for converted and methylated DNA.

FAQ - 3341