EpiTect Control DNA and Control DNA Set

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

 

 

 

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.

 

 

 

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

 

 

 

 

 

 

 

 

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.

 

 

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.