- Active Motif’s MethylCollector™ Ultra offers a fast magnetic assay capable of efficiently isolating methylated CpG islands from fragmented genomic DNA. MethylCollector Ultra is based on the Methylated CpG Island Recovery Assay (MIRA) which uses a MBD2b/MBD3L1 protein complex for improved enrichment of CpG dinucleotides. Enriched DNA can be used in many downstream applications, such as end point or real time PCR analysis of the methylation status of particular loci in normal and diseased samples, bisulfite conversion followed by cloning and sequencing, or amplification and labeling for microarray analysis.
- MethylCollector Ultra Advantages
- High affinity binding provides greater enrichment than other MBD capture or antibody-based methods
- Fast magnetic bead based protocol is completed in less than 3 hours
- Specifically detects methylated CpGs from 1 ng – 1 µg of DNA fragmented by sonication or enzymatic digestion
- Includes positive control DNA and PCR primers to ensure success
- Eluted DNA is suitable for various downstream applications
- Methylated CpG Island Recovery Assay (MIRA)
- MethylCollector™ Ultra is based on the Methylated CpG Island Recovery Assay (MIRA) which utilize a His-tagged recombinant methyl-binding protein complex, comprised of MBD2b/MBD3L1, that specifically binds methylated CpGs of genomic DNA fragments that have been prepared by sonication or enzymatic digestion. Nickel-coated magnetic beads capture the protein-DNA complexes which are separated from the rest of the genomic DNA using the included magnet. Optimized buffers ensure that fragments with little or no methylation are removed. The methylated DNA is then eluted from the beads in the presence of Proteinase K. Following clean up, the eluted DNA is ready for use in PCR analysis or other downstream applications.
- The Methylated CpG Island Recovery Assay (MIRA) utilizes the high affinity of the MBD2b/MBD3L1 complex for methylated DNA, to provide better enrichment than assays that use the methyl-binding protein MBD2 alone. MethylCollector™ Ultra can efficiently enrich for methylated DNA from as few as 170 cells (~1 ng DNA).