Quantitative Analysis of the Muscular Dystrophin Gene Using a Polymerase Chain Reaction and High-Performance Liquid Chromatography with Electrochemical Detection.

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Oligonucleotides carrying a ferrocene moiety at their 5′-end were used as a primer of polymerase chain reaction (PCR) and part of the v-myc and muscular dystrophin genes were amplified with them. The PCR products thus obtained were rapidly separated and detected with high sensitivity at femtomole (fmol) level by the HPLC-ECD system. The ECD response for the PCR products was linear over a range of 1 to 1000fmol. We tried to quantitate the muscular dystrophin gene in a chromosomal sample prepared from leukocytes of healthy men and women, which is located in their X-chromosomes. The quantitative PCR procedure with a ferrocenyl primer gave a 1.4 to 2.0 ratio of the amount of the gene for men to women, which is in reasonable agreement with the expected ratio of 1.0 to 2.0. These results demonstrate that ferrocenyl oligonucleotides serve as a useful tool for DNA analysis.
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