Comparative Analysis of Six DNA Extraction Methods in Cowpea (Vigna unguiculata L.Walp)

Huaqiang Tan, Haitao Huang, Mamman Tie, Jianyao Ma, Huanxiu Li

Abstract


High quality DNA extractions are a prerequisite for genetic studies of a variety of plants including cowpea (Vigna unguiculata). Nowadays, there are a great number of plant DNA extraction methods, and commercially available extraction kits are also becoming more and more popular. It appears that different procedures work best for different plant groups. Thus in the genetic studies of cowpea, which DNA extraction method to choose becomes a concern. To solve this problem, five classic plant DNA isolation methods, including three CTAB methods and two SDS methods, were compared and evaluated while isolation using a commercial kit was also undertaken. The DNA extracted by these six methods from two-week-old cowpea seedlings were analyzed according to their cost and time, yield, purity, integrity, and functionality in restriction endonuclease digestion and PCR (polymerase chain reaction) based downstream analysis. After the evaluation, one most suitable method, described by Dellaporta et al. (1983) was selected and chosen for isolating DNA from young leaves of cowpea seedlings. The cost and time required in this method was relatively low. In addition, the quantity and the quality of the DNA extracted by this method were high enough to perform hundreds of PCR-based reactions.


Full Text: PDF DOI: 10.5539/jas.v5n7p82

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This work is licensed under a Creative Commons Attribution 3.0 License.

Journal of Agricultural Science ISSN 1916-9752 (Print) ISSN 1916-9760 (Online)

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