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Figure 3 | BMC Molecular Biology

Figure 3

From: Genetic correction of splice site mutation in purified and enriched myoblasts isolated from mdx5cv mice

Figure 3

Quantitative measurement of dystrophin correction in primary cultures containing mdx5cv myoblasts at the DNA level and sequence analysis. A: Total genomic DNA was isolated from targeted myoblasts at various time points, 24 to 96 hours after introduction of ODNs, mdx47NT or mdx47T. Twenty-five nanograms of this DNA was subjected to absolute quantification by PCR analysis with a FAM labeled probe specific only for the wild-type dystrophin sequence. A standard curve had been previously generated using C2C12 (WT) genomic DNA and the amount of wild-type dystrophin has been extrapolated from this standard and is presented as nanograms of wild-type dystrophin. These data represent the average of three experiments that were analyzed in triplicate. Significance was measured by a Student's t-test and samples were considered significantly different with a p value of less than *0.05, **0.01, ***0.001. B: DNA sequence reaction of PCR products showing the chromatogram representing a mixed population at the targeted base (base 102) with adenine (green/corrected base) being the major peak and with thymine (red/mutant base) being a minor population within the reaction.

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