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为了建立准确、高效、实用的板栗品种鉴别体系和方法,采用改进的CTAB—DNA提取方法,从板栗的嫩叶和老叶中提取获得总DNA。所得DNA样品的D260nm/D280nm值为1.7～1.9,琼脂糖凝胶上主带清晰、较少降解、样品纯度高、蛋白去除干净且获得率高。该体系使用操作简便、快捷、高效,适合于板栗DNA提取,所提取的DNA样品不含PCR反应抑制剂及其他酶反应抑制剂,可被限制性内切酶MseI和EcoRI完全酶切,适合AFLP-PCR反应应用,得到清晰的指纹式样,适宜用作板栗品种鉴别的有效方法。
In order to establish an accurate, efficient and practical chestnut variety identification system and method, using the improved CTAB-DNA extraction method, the total DNA was extracted from young leaves and old leaves of chestnut. The obtained DNA sample had a D260nm / D280nm value of 1.7 to 1.9, the main band on the agarose gel was clear and less degraded, the purity of the sample was high, the protein was removed cleanly, and the yield was high. The system is simple to use, fast and efficient, suitable for chestnut DNA extraction. The extracted DNA samples do not contain PCR reaction inhibitors and other enzyme reaction inhibitors, and can be completely digested with restriction enzymes MseI and EcoRI, suitable for AFLP -PCR reaction to obtain a clear fingerprint pattern, suitable for chestnut varieties identification of effective methods.