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

被引:46
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
相关论文
共 7 条
[1]   Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques [J].
Aljanabi, SM ;
Martinez, I .
NUCLEIC ACIDS RESEARCH, 1997, 25 (22) :4692-4693
[2]   Biogeography and floral evolution of baobabs (Adansonia, bombacaceae) as inferred from multiple data sets [J].
Baum, DA ;
Small, RL ;
Wendel, JF .
SYSTEMATIC BIOLOGY, 1998, 47 (02) :181-207
[3]  
Doyle JJ, 1987, PHYTOCHEM B, V19, P11, DOI DOI 10.2307/4119796
[4]   Neoendemism in Madagascan scaly tree ferns results from recent, coincident diversification bursts [J].
Janssen, Thomas ;
Bystriakova, Nadia ;
Rakotondrainibe, France ;
Coomes, David ;
Labat, Jean-Noel ;
Schneider, Harald .
EVOLUTION, 2008, 62 (08) :1876-1889
[5]   A spot-PCR technique for the detection of phloem-limited grapevine viruses [J].
LaNotte, P ;
Minafra, A ;
Saldarelli, P .
JOURNAL OF VIROLOGICAL METHODS, 1997, 66 (01) :103-108
[6]   UNIVERSAL PRIMERS FOR AMPLIFICATION OF 3 NONCODING REGIONS OF CHLOROPLAST DNA [J].
TABERLET, P ;
GIELLY, L ;
PAUTOU, G ;
BOUVET, J .
PLANT MOLECULAR BIOLOGY, 1991, 17 (05) :1105-1109
[7]   THE ROLE OF VEGETATION STRUCTURE AND FUEL CHEMISTRY IN EXCLUDING FIRE FROM FOREST PATCHES IN THE FIRE-PRONE FYNBOS SHRUBLANDS OF SOUTH-AFRICA [J].
VANWILGEN, BW ;
HIGGINS, KB ;
BELLSTEDT, DU .
JOURNAL OF ECOLOGY, 1990, 78 (01) :210-222