UnMethylCollector is a novel technology that allows for positive identification ofunmethylated DNA fragments. The kit specifically enriches for unmethylated CpG islands using a CXXC binding domain from mouse Mbd1. UnMethylCollector allows researchers to identify hypomethylated promoters from normal or diseased samples. This simple and sensitive technique provides DNA suitable for use in many downstream applications, such as real time or endpoint PCR analysis of the methylation status of particular loci, sequencing, or amplification and labeling for microarray analysis.
- Fast magnetic bead based protocol is completed in less than 3 hours
- Optimized reagents and protocol ensure successful capture from DNA fragments containing only a single unmethylated CpG dinucleotide.
- Specifically detects unmethylated CpGs from 10 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
The UnMethylCollector Kit uses a His-tagged CXXC domain from mouse Mbd1 to specifically bind unmethylated CpG islands of genomic DNA fragments that have been prepared by sonication or enzymatic digestion. The kit provides two binding buffers; a low-salt buffer for use with samples containing less than 5 unmethylated CpG dinucleotides and a higher salt buffer for efficient binding of samples with a large number of unmethylated CpGs. The His-CXXC protein is added to the DNA fragments, and these protein-DNA complexes are captured with nickel-coated magnetic beads. Subsequent wash steps are performed using an optimized buffer and the included magnet to separate the unmethylated fragments from the rest of the genomic DNA. The unmethylated DNA is then eluted from the beads in the presence of high salt. Following clean up, the eluted DNA is ready for use in PCR analysis or other downstream applications.
Due to the high efficiency of UnMethylCollector and the enormous amplification capability and specificity of PCR, analysis of the methylation status of a specific genomic DNA locus can be performed on DNA isolated from less than 1600 cells (~10 ng DNA). The specificity of the CXXC domain is able to enrich for DNA fragments containing only a single unmethylated CpG.View UnMethylCollector on Active Motif’s website