EpiTect Control DNA and Control DNA Set

甲基化分析中用作实验对照

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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
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EpiTect Control DNA and Control DNA Set 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。

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Features

  • 使用便利、经严格质量控制的即用型DNA溶液
  • 亚硫酸氢盐转化的DNA,用于对照实验
  • 适用于各种甲基化分析

Product Details

EpiTect Control DNAs是即用型、完全甲基化或非甲基化经亚硫酸氢盐转化的DNA,以及未处理的非甲基化基因组DNA,为甲基化分析设立标准可靠的对照反应。甲基化或未甲基化的亚硫酸氢盐转化DNA储存在EB Buffer(10 mM Tris·Cl)中,是浓度为10 ng/μl的即用型溶液。未转化的非甲基化人类对照DNA也储存在EB Buffer(10 mM Tris·Cl)中,是浓度为50 ng/μl的即用型溶液。

Performance

EpiTect Control DNAs非常适合作为基于探针的real-time PCR甲基化分析的定量标准(参见" Real-time PCR standards") 和用于HRM甲基化分析(参见" HRM quantification standards")。
See figures

Principle

EpiTect Control DNA Set包含所有为甲基化分析设立标准可靠的对照反应的对照DNA。例如:在甲基化特异性PCR中,对照反应必须确保PCR探针和引物特异性结合甲基化或非甲基化DNA(参见" Use of EpiTect Control DNA in PCR for methylation analysis")。这些反应需要不同浓度的完全被亚硫酸氢盐转化的对照DNA,而这些DNA全部甲基化或全部非甲基化(参见" EpiTect Control DNA Pyrogram"和" EpiTYPER MALDI-TOF analysis of WBA DNA")。此外,混合的对照DNAs可做为定量标准,用于在HRM和基于探针的甲基化实验中确定甲基化水平。
See figures

Procedure

表观遗传学中标准的工作流程

表观遗传学信息不仅在生物和医学研究领域,尤其肿瘤学研究中是非常重要的,而且对干细胞研究和生物学的发展也至关重要。由于缺乏从有限样本中获得可重复数据的标准方法,分析DNA甲基化变化仍是一个难题。配合其新引进的EpiTect溶液,QIAGEN提供标准的分析前和分析溶液,用于从DNA样本收集、稳定和纯化到亚硫酸氢盐转化和real-time或终点PCR甲基化分析或测序(参见" Standardized workflows in epigenetics")。

See figures

Applications

EpiTect Control DNAs适用于各种甲基化分析的对照反应,包括:

  • 评估MSP引物的特异性
  • 作为HRM和MethyLight PCR的定量标准
  • 评估MethyLight PCR引物和探针的特异性
  • 确定亚硫酸氢盐转化反应的效率

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsEnd point Methylation Specific PCR (MSP), real-time methylation specific PCR
Number of reactionsfor 1000 control PCRs
Concentration10 ng/µl (1 µg in total) excepted the unmethylated control DNA: 50 ng/µl (10 µg in total)

Resources

试剂盒操作手册 (4)
安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
Safety Data Sheets (1)
Certificates of Analysis (1)
Kit Handbooks (4)

FAQ

What are the expected PCR results when using EpiTect Control DNA on untreated or bisulfite-converted DNA?

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
Can the EpiTect PCR Control DNAs also be used as standard in real-time methylation specific quantification?

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
Can the EpiTect PCR Control DNAs be used for mouse and rat assays?

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

 

 

 

 

 

 

 

 

FAQ ID -2008
How long can DNA converted with EpiTect Bisulfite Kits be stored before use in methylation specific PCR with the EpiTect MSP Kit?

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
3341 - How are the EpiTect DNA controls produced and tested?

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