MIRA

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.

Advantages:

  • High specificity and sensitivity, requiring as little as 1 ng input gDNA
  • Does not require DNA denaturation
  • Independent of restriction sites

Disadvantages:

  • Resolution limited to 100 bases
  • Requires a minimum of 2 methylated CpGs in the captured fragment Rauch et al., 2011)


Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

None available yet

References:

Benjamin A. L., Green B. B., Crooker B. A., McKay S. D. and Kerr D. E. Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome. BMC Genomics. 2016;17:258

Yoneda A., Henson W. R., Goldner N. K., et al. Comparative transcriptomics elucidates adaptive phenol tolerance and utilization in lipid-accumulating Rhodococcus opacus PD630. Nucleic Acids Res. 2016;44:2240-2254

Jin S. G., Xiong W., Wu X., Yang L. and Pfeifer G. P. The DNA methylation landscape of human melanoma. Genomics. 2015;106:322-330