A RAPID AND INEXPENSIVE METHOD FOR THE DIRECT PCR AMPLIFICATION OF DNA FROM PLANTS

被引:45
作者
Bellstedt, Dirk U. [1 ]
Pirie, Michael D. [1 ]
Visser, J. Christiaan [1 ]
de Villiers, Margaret J. [1 ]
Gehrke, Berit [2 ]
机构
[1] Univ Stellenbosch, Dept Biochem, ZA-7600 Stellenbosch, South Africa
[2] Univ Cape Town, Dept Bot, ZA-7700 Rondebosch, South Africa
基金
新加坡国家研究基金会;
关键词
Direct PCR; DNA isolation; plant biotechnology; plant systematics; UNIVERSAL;
D O I
10.3732/ajb.1000181
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Premise of the study: We present a rapid and inexpensive alternative to DNA isolation for polymerase chain reaction (PCR) amplification from plants. Methods and Results: The method involves direct PCR amplification from material macerated in one buffer, followed by dilution and incubation in a second buffer. We describe the procedure and demonstrate its application for nuclear and plastid DNA amplification across a broad range of vascular plants. Conclusions: The method is fast, easy to perform, cost-effective, and consequently ideal for large sample numbers. It represents a considerable simplification of present approaches requiring DNA isolation prior to PCR amplification and will be useful in plant systematics and biotechnology, including applications such as DNA barcoding.
引用
收藏
页码:E65 / E68
页数:4
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