Understanding DNA methylation is critical for learning about control of transcription, genomic imprinting, embryonic development, and carcinogenesis. Diagenode proposes two main approaches to studying DNA methylation - bisulfite conversion with downstream sequencing or qPCR and immunoprecipitation (IP).
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 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.
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.
| Cat. No. | Product | Format | Price | |
|---|---|---|---|---|
| C02030030 |
Premium Bisulfite kit Make your Bisulfite conversion now in only 60 minutes ! Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion... |
50 rxns | $240.00 | |
| C02030036 |
Premium RRBS kit V2 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 t... |
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 together with their corresponding qPCR primer sets th... |
48 rxns | $275.00 | |
| C09010010 |
MethylTaq DNA polymerase MethylTaq DNA polymerase is a high-performance Hot Start thermostable recombinant DNA polymerase. MethylTaq is an extremely robust modified Taq DNA polymeras... |
250 units | $260.00 | |
| View more products from the Bisulfite conversion product category... | ||||
| Cat. No. | Product | Format | Price | |
|---|---|---|---|---|
| C02010021 |
MagMeDIP Kit Perform MeDIP (Methylated DNA Immunoprecipitation) followed by qPCR or NGS to estimate DNA methylation status of your sample using a highly sensitive 5-met... |
48 rxns (IP) | $750.00 | |
| C02010014 |
Auto MagMeDIP qPCR Kit - ordering reference: C02010021 The reference C02010014 has been replaced by C02010021. Perform MeDIP (Methylated DNA Immunoprecipitation) on the SX-8G IP-S... |
48 rxns | ||
| C02040012 |
DNA methylation control package The DNA methylation control package includes one methylated and one unmethylated spike-in controls together with their corresponding qPCR primer ... |
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 together with their corresponding qPCR primer sets th... |
48 rxns | $275.00 |
| Cat. No. | Product | Format | Price | |
|---|---|---|---|---|
| C02020010 |
MethylCap kit The MethylCap kit allows to specifically capture DNA fragments containing methylated CpGs. The assay is based on the affinity purification of methylated DNA ... |
48 rxns | ||
| C02020011 |
Auto MethylCap kit The Auto MethylCap kit allows to specifically capture DNA fragments containing methylated CpGs. The assay is based on the affinity purification of methylated... |
48 rxns | ||
| C02020012 |
MethylCap protein The MethylCap protein has been extensively validated for specific isolation of DNA fragments containing methylated CpGs. It consists of the methyl-CpG-bindin... |
100 µg |
| Cat. No. | Product | Format | Price | |
|---|---|---|---|---|
| C02010031 |
hMeDIP kit x16 (monoclonal mouse antibody) The hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented genomic DNA samples for use in genome-wide methylation analysi... |
16 rxns | $690.00 | |
| C02010034 |
Auto hMeDIP kit x16 (monoclonal mouse antibody) The Auro hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented genomic DNA samples for use in genome-wide methylation anal... |
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 set that can be added to the DNA... |
48 rxns | $150.00 |
| Cat. No. | Product | Format | Price | |
|---|---|---|---|---|
| C02040018 |
DNA hydroxymethylation control package The DNA hydroxymethylation control package includes one hydroxymethylated spike-in control and its corresponding qPCR primer set that can be added to the DNA... |
48 rxns | $150.00 | |
| C02040012 |
DNA methylation control package The DNA methylation control package includes one methylated and one unmethylated spike-in controls together with their corresponding qPCR primer ... |
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 together with their corresponding qPCR primer sets th... |
48 rxns | $275.00 |
| Cat. No. | Product | Format | Price | |
|---|---|---|---|---|
| C09010010 |
MethylTaq DNA polymerase MethylTaq DNA polymerase is a high-performance Hot Start thermostable recombinant DNA polymerase. MethylTaq is an extremely robust modified Taq DNA polymeras... |
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 bisulfite converted NGS libraries, e... |
135 μL | $120.00 |
