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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Product
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Cat. no.
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List price:
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EpiTect Control DNA, methylated (100)
Methylated and bisulfite converted human control DNA for 100 control PCRs
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59655
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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
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59695
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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.
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).| |
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
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Feature
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Specifications
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Applications
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End point Methylation Specific PCR (MSP), real-time methylation specific PCR
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Concentration
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10 ng/µl (1 µg in total) excepted the unmethylated control DNA: 50 ng/µl (10 µg in total)
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Number of reactions
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for 1000 control PCRs
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Download Safety Data Sheets for QIAGEN product components.
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View
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Images
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.
HRM quantification standards.
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.
EpiTect Control DNA Pyrogram.
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).
Use of EpiTect Control DNA in PCR for methylation analysis.
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.
EpiTYPER MALDI-TOF analysis of WBA 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).
Standardized workflows in epigenetics.
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