Development of DNA point mutation detection methods using probe DNA-immobilized microbeads

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  • DNAプローブ固定化微粒子を用いたDNA一塩基変異検出法の開発
  • DNA プローブ コテイカ ビリュウシ オ モチイタ DNA イチエンキ ヘンイ ケンシュツホウ ノ カイハツ

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Abstract

One technique that can distinguish low-abundant mutant DNA from wild-type DNA is the ligase detection reaction (LDR) coupled to a primary polymerase chain reaction (PCR). The LDR products as a target obtained by the sequential PCR/LDR reactions can be analyzed in a variety of fashions such as microarray and slab gel electrophoresis. In the present work, we employed probe DNA-immobilized magnetic microbeads to selectively capture the fluorescently labeled-LDR products via the target hybridization with the probe. After the hybridization, the microbeads were rinsed and introduced into a capillary tube and packed in one space in the tube with a magnet. Photoluminescence from the beads aggregates in the tube was successfully obtained with fluorescence or chemiluminescence detection system, showing that the present method can be amenable to mutation detection.

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