A novel methylation PCR that offers standardized determination of FMR1 methylation and CGG repeat length without southern blot analysis.

ABSTRACT: Fragile X syndrome and associated disorders are characterized by the number of CGG repeats and methylation status of the FMR1 gene for which Southern blot (SB) historically has been required for analysis. This study describes a simple PCR-only workflow (mPCR) to replace SB analysis, that incorporates novel procedural controls, treatment of the DNA in separate control and methylation-sensitive restriction endonuclease reactions, amplification with labeled primers, and two-color amplicon sizing by capillary electrophoresis. mPCR was evaluated in two independent laboratories with 76 residual clinical samples that represented typical and challenging fragile X alleles in both males and females. mPCR enabled superior size resolution and analytical sensitivity for size and methylation mosaicism c

SUBMITTER: Grasso M 

PROVIDER: S-EPMC3873488 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature