EpiTect Control DNA and Control DNA Set

甲基化分析中用作实验对照
  • 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

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。

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EpiTect Control DNA, methylated (100)
Methylated and bisulfite converted human control DNA for 100 control PCRs
59655
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EpiTect Control DNA, unmethlyated (100)
Unmethylated and bisulfite converted human control DNA for 100 control PCRs
59665
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EpiTect PCR Control DNA Set (100)
Human control DNA set (containing both bisulfite converted methylated and unmethylated DNA and unconverted unmethylated DNA) for 100 control PCRs
59695
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EpiTect Control DNA (1000)
Unmethylated human control DNA for 1000 control PCRs
59568
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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.


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Real-time PCR standards.|HRM quantification standards.|EpiTect Control DNA Pyrogram.|Use of EpiTect Control DNA in PCR for methylation analysis.|EpiTYPER MALDI-TOF analysis of WBA DNA.|Standardized workflows in epigenetics.|
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.|A CpG island from the promoter region of the APC gene (adenomatosis polyposis coli) was amplified, and the degree of methylation was determined by HRM methylation analysis on the Rotor-Gene Q, using the EpiTect HRM PCR Kit. Methylated and unmethylated DNA from the EpiTect Control DNA Set was mixed in varying ratios, and used as the templates for the analysis.
|Sixteen CpGs of the MGMT promoter were checked for complete methylation (mean methylation rate >95%) or complete nonmethylation (mean methylation rate 0.3%). The PCR products for MGMT (6-O methylguanine-DNA methyltransferase), which is a p53-related gene involved in DNA repair and drug resistance, were subjected to Pyrosequencing, which was performed on a PyroMark Q96ID instrument. (Data kindly provided by Uwe Gerstenmaier, Varionostic GmbH, Ulm).|The methylation-specific primer (M-Primer) will only anneal to the control DNA that is fully methylated and bisulfite converted. The primer specific for unmethylated DNA (U-Primer) will only anneal to the control DNA that is fully unmethylated and bisulfite converted. Neither M-Primer nor U-Primer will anneal to fully unmethylated genomic DNA.|Methylation of the MP6 locus was measured in unmethylated DNA (UMDNA 1), methylated DNA (VIAL A 1), and a mixture of both (HTXA 1), before and after amplification (WBAUMDNA1, WBAHTXA 1, WBAVIAL A 1) using the EpiTect Whole Bisulfitome Kit. The methylation pattern prior to amplification is very similar to that of the amplified DNA, demonstrating representative amplification. (Data kindly provided by Hany Ezzeldin, Mayo Clinic, Rochester, USA).|  |
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")。 
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。 
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")。

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
Feature
Specifications
Applications End point Methylation Specific PCR (MSP), real-time methylation specific PCR
Concentration 10 ng/µl (1 µg in total) excepted the unmethylated control DNA: 50 ng/µl (10 µg in total)
Number of reactions for 1000 control PCRs

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