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Methylation-Specific Multiplex Ligation-Dependent Probe Amplification Enables a Rapid and Reliable Distinction between Male FMR1 Premutation and Full-Mutation.

Published byVera Tanuwidjaja Modified over 5 years ago

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Presentation on theme: "Methylation-Specific Multiplex Ligation-Dependent Probe Amplification Enables a Rapid and Reliable Distinction between Male FMR1 Premutation and Full-Mutation."— Presentation transcript:

1 Methylation-Specific Multiplex Ligation-Dependent Probe Amplification Enables a Rapid and Reliable Distinction between Male FMR1 Premutation and Full-Mutation Alleles  Anders O.H. Nygren, Sylvia I. Lens, Ralph Carvalho  The Journal of Molecular Diagnostics  Volume 10, Issue 6, Pages (November 2008) DOI: /jmoldx Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 MS-MLPA principle: MS-MLPA probes hybridize in juxtaposition to completely denatured sample DNA. On completion of probe hybridization, ligation of the MS-MLPA probe oligonucleotides is combined with digestion of the probe/sample DNA hybrid complex using the methylation-sensitive restriction endonuclease HhaI. After the enzymatic digestion, probes juxtaposed to methylated regions will generate a signal in the subsequent PCR, whereas probes adjacent to unmethylated sites will be degraded and will generate no signal. The Journal of Molecular Diagnostics  , DOI: ( /jmoldx ) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 Electropherograms depicting the differences in FMR1 and FMR2 peaks in each of the three male patient groups: Promega control (A and B), premutation patient (C and D), and full-mutation patient (E and F). All five FMR1 peaks are missing in both the Promega control and the premutation HhaI-digested samples (B and D) and present in the full-mutation HhaI-digested sample (F), indicative of FMR1 promoter hypermethylation only in the full-mutation patient; all five FMR2 peaks are missing from the HhaI-digested samples of all three groups (B, D, and F), indicative of lack of FMR2 promoter hypermethylation; all three control peaks showed complete digestion. Straight arrows, methylation-specific FMR1 probes; curved arrows, methylation-specific FMR2 probes; arrowheads, digestion control probes. The Journal of Molecular Diagnostics  , DOI: ( /jmoldx ) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

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