ホルマリン固定組織からのDNA抽出法とPCR法による遺伝子解析

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タイトル別名
  • DNA extraction and sequencing using PCR from formalin-fixed samples
  • ホルマリン コテイ ソシキ カラ ノ DNA チュウシュツホウ ト PCRホウ ニ ヨル イデンシ カイセキ

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Formalin is a main fixative in the field of pathology. Molecular biological analysis of formalin-fixed samples was difficult because formalin fixation decreased the quality of isolated DNA. Therefore, we compared the quality of DNA obtained by using DNA extraction kit （Sepa GeneR） to that using proteinase K. Using proteinase K, it was possible to extract high quality DNA, and obtain DNA from samples of 3 months fixative. Moreover, by proteinase K method, it was also possible to analyze aprataxin gene exon 5 in DNA extraction from formalin-fixed human brain tissues from a suspected case of early-onset ataxia with ocular motor apraxia and hypoalbuminemia （EAOH）. The aprataxin gene exon 5 DNA sequences were obtained following in vitro gene amplification using nested-PCR. Mutation on aprataxin gene exon5 was not observed in the suspected case of EAOH; however, it was possible to perform sequence analysis of aprataxin gene exon5. This method was more useful for DNA extraction and direct sequencing of formalin-fixation samples than the kit method.

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