Methylamp Methylated DNA Capture Kit (Enrichment of Methylated DNA from Genomic DNA)

Product Description

Please see the EpiQuik™ Methylated DNA Immunoprecipitation Kit if you are working with nuclear extract as the starting material.

The Methylamp™ Methylated DNA Capture (MeDIP) Kit is a complete set of optimized reagents to enrich and capture methylated DNA fragments in a convenient microplate-based format. The method, methylated DNA immunoprecipitation (meDIP), uses a monoclonal antibody specific to 5-methylcytosine to immunoprecipitate methylated genomic DNA. The enriched methylated fractions can then be used for gene-specific DNA methylation analysis on a genome wide scale. The Methylamp™ Methylated DNA Capture Kit is suitable for combining the specificity of enriched methylated DNA with qualitative and quantitative PCR, traditional sequencing and next-generation sequencing, DNA microarrays, as well as Southern blot.  The kit has the following advantages and features:

• Highly efficient enrichment of methylated DNA: > 98%.
• The fastest procedure available, which can be finished within 3 hours.
• Strip microplate format makes the assay flexible: manual or high throughput.
• Columns for DNA purification are included: save time and reduce labor.
• Compatible with all DNA amplification-based approaches.
• Simple, reliable, and consistent assay conditions.

Background Information

MeDIP is a genome-wide, large scale technique to enrich methylated DNA by isolating methylated DNA fragments with a 5-methylcytosine antibody. This technique has played an important role in helping to identify abnormally methylated genes in diseases such as cancer. With MeDIP, the immunoprecipitated fractions of methylated DNA can be determined by PCR to evaluate the methylation state of individual regions. Additionally, the purified fractions can be input to high throughput DNA detection methods such as DNA microarrays (MeDIP-chip) and next-generation sequencing (MeDIP-seq), becoming a useful approach for methylome-level analysis and for developing comprehensive profiles of DNA methylation on a genome-wide scale. 

Principle & Procedure

The most common and crucial experimental factor of MeDIP that may affect its accuracy involves the quality and cross-reactivity of the 5-methylcytosine antibody used in the procedure. Epigentek overcame such a limitation through extensive research and development to create a highly efficient MeDIP kit by applying a non-cross-reactive 5mC antibody. In the kit’s assay, DNA is sheared and then added into a microplate well or multiple wells in a high throughput format. A high quality ChIP/MeDIP-grade antibody specific to methylcytosine is then used to capture methylated DNA in the wells. The antibody-captured methylated DNA is released and is finally purified through the included spin columns. The eluted methylated DNA can now be used in various down-stream applications.

Performance Data

Fig.2. For enrichment of methylated DNA using the kit, DNA (0.5 ug) isolated from MCF-7 cells was added into the microwell. Methylated DNA was captured by 5-mC antibody prebound to the microwells.

Fig.3. Captured methylated DNA was used for analyzing methylation level of GAPDH and MLH1 promoter with the use of primers and probes specific to GAPDH and MLH1 promoters, respectively.