A complete set of optimized buffers and reagents to carry out a successful hMeDIP procedure for the selective enrichment of DNA fragments containing 5-hydroxymethylcytosine (5-hmC) in a high throughput format using DNA isolated from any species.
The hydroxymethylated DNA that is enriched with this kit can be used in various downstream applications including PCR (hMeDIP-PCR) and microarrays (hMeDIP-chip) for investigating gene-specific hydroxymethylation in cells or tissues.
Because of the presence of 5-hmC in DNA with unclear functions in gene regulation and because of the discovery of enzymes that produce 5-hmC, it is crucial to identify hydroxymethylation status in specific gene loci, which would help to better understand methylation-based epigenetic regulation of gene functions. To achieve this, Epigentek has developed an innovative method to capture DNA fragments containing 5-hmC, and incorporated this method into its EpiQuik Hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit. This kit uses a high affinity 5-hmC antibody to selectively capture double-stranded or single stranded DNA fragments containing 5-hmC.
The kit has the following features:
• Extremely fast and convenient protocol with a total procedure time (from input sample to ready-to-use hydroxymethylated DNA) of less than 3 hours, which includes a minimal handling time of less than 20 minutes.
• Flexible 96 stripwell microplate format makes the assay very easy to handle: manual method with one reaction at a time or high throughput method with 96 reactions at a time.
• Highly efficient enrichment ratio of positive/negative control > 1000.
• Low DNA input requirement of as low as 0.1 ug per reaction.
• High reproducibility through pre-optimized hMeDIP conditions.
• Compatible with various downstream analysis workflows including hMeDIP-PCR and hMeDIP-chip.