Simple and quick methods for nematode DNA preparation
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A DNA extraction kit, ISOHAIR® (Nippon Gene), which was originally developed for preparing DNA from hair and nail samples, was used to prepare nematode DNA for PCR and sequencing analyses. The methods provided here, which involved digesting (resolving) a single nematode individual in a tube containing the mixed enzyme solution, enabled the DNA to be prepared within 20 min. The prepared DNA was suitable for PCR amplification of near-full-length small subunit ribosomal RNA (ca 1.7 kb), of the D2/D3 expansion segments of large subunit RNA (ca. 0.7 kb), and of partial mitochondrial COI (ca. 0.7 kb) genes, followed by sequencing analysis. Furthermore, the prepared material could be preserved in a freezer (−30 °C) for at least 20 months, and more than 300 PCR reactions could be performed from a single individual nematode.
KeywordsDNA ISOHAIR® Nematode PCR Sequencing
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