DNAメチル化についての理解は、転写の制御、ゲノムインプリンティング、胚発生、および発癌の研究に不可欠です。 Diagenodeは、シーケンシングまたはqPCRによるバイサルファイト変換と免疫沈降（IP）を提案しており、これらはDNAメチル化研究の2つの主要なアプローチです。
Generate bisulfite converted gDNA libraries ready for gDNA single or paired-end sequencing of the whole genome
Generate bisulfite converted libraries ready for single and paired-end sequencing of CpG-rich parts of the genome
OR
Bisulfite convert gDNA ready for region-specific (q)PCR
Precipitate hydroxy-/methylated DNA with a specific antibody
Capture methylated DNA via methyl-binding domain (MBD) of human MeCP2 protein
Bisulfite modification of DNA is the most commonly used, "gold standard" method for DNA methylation studies providing single nucleotide resolution. This technology is based on the chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at the singe nucleotide level. Various analyses can be performed on the altered sequence to retrieve this information: bisulfite sequencing, pyrosequencing, methylation-specific PCR, high resolution melting curve analysis, microarray-based approaches, and next-generation sequencing.
Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected (see Figures).
The MBD technology used in our MethylCap Kit is based on the very high affinity of a H6-GST-MBD fusion protein for methylated DNA. This protein consists of the methyl binding domain (MBD) of human MeCP2, as a C-terminal fusion with Glutathione-S-transferase (GST) containing an N-terminal His6-tag. The H6-GST-MBD fusion protein can be used to specifically isolate DNA containing methylated CpGs.
Diagenode’s MethylCap Kit enables high enrichment of double-stranded DNA and a differential fractionation in function of the methylated CpG density. Fractionation reduces the complexity of samples and makes subsequent next generation sequencing easier. Prior to the MethylCap assay, DNA is first extracted and sheared using the Bioruptor® Sonicator.
The Methylated DNA IP (MeDIP) is based on the affinity purification of methylated DNA using an antibody directed against 5-methylcytosine (5-mC) or 5-hydroxymethylcytosine (5-hmC) in the case of hMeDIP.
In brief, Methyl DNA IP is performed as follows: Genomic DNA from cultured cells or tissues is prepared, sheared, and then denatured. Then, immunoselection and immunoprecipitation can take place using the antibody directed against 5 methylcytosine and antibody binding beads. After isolation and purification is performed, the IP’d methylated DNA is ready for any subsequent analysis as qPCR, amplification, hybridization on microarrays or next generation sequencing.
| Cat. No. | 製品 | Format | Price | |
|---|---|---|---|---|
| C02030030 |
Premium Bisulfite kit Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate... |
50 rxns | $240.00 | |
| C02030036 |
Premium RRBS kit V2 (Coming soon) Reduced Representation Bisulfite Sequencing (RRBS) offers a cost-effective, targeted solution to perform genome-scale DNA methylation analysis... |
24 rxns | $1,590.00 | |
| C02030037 |
Premium RRBS kit V2 x96 RRBS for low DNA amounts and accurate analysis Reduced Representation Bisulfite Sequencing (RRBS) offers a cost-effective, focused solution to perform genome-scale DNA methylation analysis at the s... |
96 rxns | $3,745.00 | |
| C02030031 |
Auto Premium Bisulfite kit Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate... |
40 rxns | $240.00 | |
| C02030035 |
Bisulfite conversion reagent for RRBS This reagent is compatible with the Premium RRBS Kit and provides reagents for additional bisulfite conversion reactions. Reduced Representation Bisulfite S... |
8 rxns | $120.00 | |
| C02040019 |
DNA methylation control package V2 The DNA methylation control package V2 includes one methylated and one unmethylated spike-in controls that can be added directly to the DNA sample of interes... |
48 rxns | $275.00 | |
| C09010010 |
MethylTaq DNA polymerase MethylTaq DNAポリメラーゼは、高性能ホットスタート熱安定性組換え型DNAポリメラーゼです。 MethylTaqは、74℃以下のすべての活性を完全に欠如しているため、低温での非特異的プライミングを回避する、非常に頑強に修飾されたTaq DNAポリメラーゼです。 この強力な酵素は、要求の厳しい用途でも優れ... |
250 units | $260.00 | |
| View more products from the バイサルファイト変換 product category... | ||||
| Cat. No. | 製品 | Format | Price | |
|---|---|---|---|---|
| C02010021 |
MagMeDIP qPCR kit Sensitive tumour detection and classification using plasma cell-free DNA methylomesRead the publication Preparation of cfMeDIP-seq libraries for methylo... |
48 rxns (IP) | $750.00 | |
| C02010014 |
Auto MagMeDIP kit Perform Automated MeDIP (seq) (Methylated DNA Immunoprecipitation sequencing) with Diagenode´s Auto MagMeDIP kit to estimate DNA methylation status of ... |
48 rxns | ||
| C02040012 |
DNA Methylation control package The DNA Methylation control package includes unmethylated and in vitro methylated DNA together with specific primer sets for assessing the efficiency of your... |
40 rxns | $300.00 | |
| C02040019 |
DNA methylation control package V2 The DNA methylation control package V2 includes one methylated and one unmethylated spike-in controls that can be added directly to the DNA sample of interes... |
48 rxns | $275.00 |
| Cat. No. | 製品 | Format | Price | |
|---|---|---|---|---|
| C02020010 |
MethylCap kit MethylCapキットを用いたアッセイは、ヒトMeCP2タンパク質のメチルCpG結合ドメイン（MBD）を使用したメチル化DNAのアフィニティー精製に基づき、メチル化されたCpGを含むDNAフラグメントを特異的にキャプチャーします。手順は手動プロセスまたはIP-Star®コンパクト自動システムの両方で行え... |
48 rxns | ||
| C02020011 |
Auto MethylCap kit The MethylCap assay is based on the affinity purification of methylated DNA using a MBD (Methyl binding domain) protein . The Diagenode’s Auto MethylCa... |
48 rxns | $695.00 | |
| C02020012 |
MethylCap protein メチル化CpGを含むDNAを特異的に単離するには、H6-GST-MBD融合タンパク質を用います。このたんぱく質はN末端His6タグを含むグルタチオン-S-トランスフェラーゼ（GST）とのC末端融合物として、ヒトMeCP2のメチル結合ドメイン（MBD）から構成されています。*このキットで溶解およびせん断することがで... |
100 µg | $565.00 |
| Cat. No. | 製品 | Format | Price | |
|---|---|---|---|---|
| C02010031 |
hMeDIP kit x16 (monoclonal mouse antibody) このキットは、5-ヒドロキシメチルシトシン（5-hmC）(20µl, 50µl, 100µl)に対する最初で唯一のモノクローナル抗体を用いたDNA免疫沈降実験専用に設計されています。キットには、免疫沈降の効率を評価するコントロール配列（5-hmC、5-mCお... |
16 rxns | $690.00 | |
| C02010034 |
Auto hMeDIP kit x16 (monoclonal mouse antibody) This kit has been designed to perform Hydroxymethylated DNA Immunoprecipitation using an antibody against 5-hydroxymethylcytosine (5-hmC) on our Automated Sy... |
16 rxns | $690.00 | |
| C02040018 |
DNA hydroxymethylation control package The DNA hydroxymethylation control package includes one hydroxymethylated spike-in control and its corresponding qPCR primer pair. The control can be ad... |
48 rxns | $150.00 |
| Cat. No. | 製品 | Format | Price | |
|---|---|---|---|---|
| C02040018 |
DNA hydroxymethylation control package The DNA hydroxymethylation control package includes one hydroxymethylated spike-in control and its corresponding qPCR primer pair. The control can be ad... |
48 rxns | $150.00 | |
| C02040012 |
DNA Methylation control package The DNA Methylation control package includes unmethylated and in vitro methylated DNA together with specific primer sets for assessing the efficiency of your... |
40 rxns | $300.00 | |
| C02040019 |
DNA methylation control package V2 The DNA methylation control package V2 includes one methylated and one unmethylated spike-in controls that can be added directly to the DNA sample of interes... |
48 rxns | $275.00 |
| Cat. No. | 製品 | Format | Price | |
|---|---|---|---|---|
| C09010010 |
MethylTaq DNA polymerase MethylTaq DNAポリメラーゼは、高性能ホットスタート熱安定性組換え型DNAポリメラーゼです。 MethylTaqは、74℃以下のすべての活性を完全に欠如しているため、低温での非特異的プライミングを回避する、非常に頑強に修飾されたTaq DNAポリメラーゼです。 この強力な酵素は、要求の厳しい用途でも優れ... |
250 units | $260.00 | |
| C09010012 |
MethylTaq Plus 2X Master Mix MethylTaq Plus 2X Master Mix is a ready-to-use cocktail containing all components required for the amplification of bisulfiteconvertedNGS libraries, except p... |
135 μL | $120.00 |
