EpiTect Control DNA and Control DNA Set

メチル化解析におけるコントロール実験
  • 即使用可能な品質管理されたDNA溶液による簡便化
  • コントロール実験用のBisulfite変換済みのDNA
  • 全てのDNAメチル化解析に最適

EpiTect Control DNA は、即使用可能なBisulfite 変換した完全メチル化DNA、Bisulfite 変換した非メチル化DNA およびBisulfite 変換していない非メチル化ゲノムDNA から構成され、メチル化解析の標準化のための信頼できるコントロール反応が可能になります。Bisulfite変換済みのメチル化/非メチル化DNA溶液はEB Buffer(10 mM Tris·Cl)で10 ng/µlに溶解されており、すぐに使用できます。Bisulfite変換していない非メチル化ヒトコントロールDNA 溶液はEB Buffer (10 mM Tris·Cl)で50 ng/μl に溶解されており、すぐに使用できます。

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EpiTect Control DNA, methylated (100)
Methylated and bisulfite converted human control DNA for 100 control PCRs
59655
€ 197,00
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EpiTect Control DNA, unmethlyated (100)
Unmethylated and bisulfite converted human control DNA for 100 control PCRs
59665
€ 197,00
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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
€ 395,00
Add to cart
EpiTect Control DNA (1000)
Unmethylated human control DNA for 1000 control PCRs
59568
€ 197,00
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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は、プローブベースのリアルタイムPCRによるメチル化解析の定量スタンダード(図 "リアルタイムPCRスタンダード")やHRM(high-resolution melting)法によるメチル化解析の定量スタンダード(図 "HRM定量スタンダード")として最適です。
Principle
The EpiTect Control DNA Setは、信頼性の高い標準化メチル化コントロール反応に必要なコントロールDNAを全て含んでおり、即使用可能なキットフォーマットになっています。例えば、MSP(メチル化特異的PCR)では、PCRプローブとプライマーがメチル化DNAまたは非メチル化DNAに特異的に結合していることを確認するためにコントロール反応を行なう必要があります(図 " MSP反応にはEpiTect Control DNA")。そのような反応では、完全にメチル化または非メチル化であるDNAを完全にBisulfite変換したコントロールDNAを様々な濃度に調製する必要があります(図 "EpiTect Control DNAのパイログラム" および "Bisulfite変換および全Bisulfitome増幅 (WBA) DNAのEpiTYPER® MALDI-TOF 解析")。また、このようなコントロールDNAを混合したものは、HRM法またはプローブによるメチル化解析でメチル化の程度を決定する際に定量スタンダードとして使用できます。
Procedure
エピジェネティクスにおける標準化されたワークフロー

エピジェネティクス情報を得ることは生物学や医学研究における多くの分野(特に、腫瘍学や幹細胞研究、および発生生物学)にとって最も重要です。しかし、DNA メチル化における変化の解析は、サンプル(特に非常に限られた量のサンプル)から再現性のあるデータを得るための標準化された方法がないために非常に困難です。QIAGENが新しく紹介するEpiTectソリューションにより、DNAサンプル採取、安定化、精製からBisulfite処理、リアルタイムPCRやエンドポイントPCR、メチル化解析、シークエンシングまで、標準化されたプレアナリティカル用製品および解析用製品を提供致します。(図 "エピジェネティクスにおける標準化されたワークフロー")。

Applications

EpiTect Control DNAは以下のような全てのメチル化解析におけるコントロール反応として最適です。

MSP用プライマーの特異性の評価
HRMやMethyLight PCRにおける定量スタンダード
MethyLight PCR用プライマーやプローブの特異性の評価
Bisulfite変換反応効率のチェック
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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Images
Standards for quantitative, real-time PCR analysis
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.

Standard curve for the quantification of the methylation degree in HRM analysis
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.
Typical Pyrograms Obtained for Unmethylated and Methylated EpiTect
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
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 Bisulfite Converted and Whole Bisulfitome Amplified (WBA)
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
Standardized workflows in epigenetics.