Diagenode

DNA methylation

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).

Diagenode Product Solutions

Diagenode Technologies

  • Bisulfite conversion


    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.

    How it works

    Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected (see Figures).

    ADVANTAGES

    • Single nucleotide resolution
    • Gene-specific and genome-wide analyses
    • NGS compatible

    Downstream analysis techniques

  • Methylated DNA immunoprecipitation

    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.

    How it works

    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.

    Overview


        QPCR

        NGS

    ADVANTAGES

    • Unaffected DNA
    • High enrichment yield
    • Robust & reproducible techniques
    • NGS compatible

    Find the solution you need

    WHOLE GENOME BISULFITE-SEQ
    REDUCE REPRESENTATION BISULFITE-SEQ
    BISULFITE CONVERSION WITH QPCR, SANGER OR PYROS
    IMMUNOPRECIPITATION WITH ANTIBODIES
    IMMUNOPRECIPITATION WITH METHYL-BINDING DOMAIN
  • Methylbinding domain protein

    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.

    ADVANTAGES

    • Unaffected DNA
    • Robust & reproducible technique
    • NGS compatible

Bisulfite conversion

Cat. No.ProductFormatPrice
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...

Methylated DNA immunoprecipitation

Cat. No.ProductFormatPrice
C02010021 MagMeDIP Kit
  Sensitive tumour detection and classification using plasma cell-free DNA methylomesRead the publication Preparation of cfMeDIP-seq libraries f...
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

Methylbinding domain protein

Cat. No.ProductFormatPrice
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 $695.00
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 $695.00
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 $565.00

Hydroxymethylated DNA immunoprecipitation

Cat. No.ProductFormatPrice
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

DNA methylation standard

Cat. No.ProductFormatPrice
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

PCR enzymes and nucleotides

Cat. No.ProductFormatPrice
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

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