荒漠植物白刺总DNA提取及鉴定

摘要/Abstract

摘要： 荒漠植物白刺含有较多的多糖、酚类等次生代谢物，用常规的CTAB、SDS等方法难以获得高质量的总DNA。针对这一问题探索出一种适合白刺属植物总DNA提取的方法。其特点是在裂解细胞膜之前，首先用无CTAB缓冲液进行两次多糖、酚类等次生代谢物的抽提；加大CTAB浓度，用3×CTAB作为裂解液；在用异丙醇沉淀DNA之前，用高浓度NaCl再次分离多糖。用这种方法我们对分布于甘肃的四种白刺属植物的总DNA进行了提取，开展了与总DNA相关的PCR扩增和其他遗传学分析，并取得满意的结果。

关键词: 白刺, 总DNA制备, trnL-F

Abstract: The content of polysaccharides, phenolic compound and other secondary metabolites is high in desert plants of Nitraria, which influences strongly the yield and quality of total DNA when being extracted by routine methods of CTAB and SDS. In this study, a new method was introduced which do best for the purity and concentration of total DNA. The characteristic of this method is as follows: to wash out polysaccharides, phenolic compound and other secondary metabolites by free-CTAB buffer twice before extracting DNA; adding the concentration of CTAB, adoption 3% CTAB rather than 2% CTAB; using high concentration of NaCl prior to DNA precipitation with isopropanol to remove residual polysaccharides. By this method, total DNA of four species of Nitraria was extracted. The result showed that it could effectively eliminate the affection of secondary materials to extracted total DNA from Nitraria, and which was suitable for PCR of chloroplast trnL-F.

Key words: Nitraria ssp., total DNA extraction, trn L-F

中图分类号:

Q943

张勇;;李鸣;杨同文;张腾国;陈拓;安黎哲;*. 荒漠植物白刺总DNA提取及鉴定[J]. 中国沙漠, 2006, 26(3): 489-492.

ZHANG Yong; LI Ming; YANG Tong-wen;ZHANG Teng-guo;CHENG Tuo;AN Li-zhe;. Preparation of Total DNA from Desert Plant Nitraria L.[J]. JOURNAL OF DESERT RESEARCH, 2006, 26(3): 489-492.

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