Methylated-CpG Island Recovery Assay (MIRA)
MIRA uses the high affinity of a methylated-CpG-binding protein complex (MBD2B and MBD3L1) to enrich regions with methylated CpG dinucleotides (Rauch et al., 2010). This approach can be applied to both array-based DNA analysis and NGS, which are sometimes distinguished as MIRA-chip (Jung et al., 2015) and MIRA-seq (Rauch et al., 2010).
In this procedure, the fragmented gDNA is incubated with purified GST-MBD2B and His-MBD3L1 proteins. The high-affinity MBD2B/MBD3L1 complex binds to the methylated gDNA templates. The methylated gDNA fragments are captured on glutathione-coated magnetic beads. The enriched methylated DNA fraction is amplified, labeled, and analyzed by NGS.
- High specificity and sensitivity, requiring as little as 1 ng input gDNA
- Does not require DNA denaturation
- Independent of restriction sites
- Resolution limited to 100 bases
- Requires a minimum of 2 methylated CpGs in the captured fragment Rauch et al., 2011)
Illumina Library prep and Array Kit Selector
None available yet
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