A Method for Isolation and Purification of Peanut Genomic DNA Suitable for Analytical Applications

被引：31

作者：

Sharma K.K. ^{[1
]}

Lavanya M. ^{[1
]}

Anjaiah V. ^{[1
]}

机构：

[1] Genetic Transformation Laboratory, Genetic Rsrc. and Enhancement Prog., Intl. Crops Res. Inst. Semi-Arid T., Patancheru

来源：

Plant Molecular Biology Reporter | 2000年 / 18卷 / 4期

关键词：

DNA isolation; PCR amplification; Peanut; Southern hybridization; Transgenic plants;

D O I：

10.1007/BF02825068

中图分类号：

学科分类号：

摘要：

Numerous methods are available for isolation of plant genomic DNA, but in practice these procedures are empirical due to variability in plant tissue composition. Consistent isolation of quality DNA from peanut (Arachis hypogaea L.) is particularly problematic due to the presence of phenolic compounds and polysaccharides. Inconsistencies in extraction results can be attributed to the age and growth stage of the plant material analyzed. Mature leaves have higher quantities of polyphenols, tannins, and polysaccharides that can contaminate DNA during isolation. We show that four published protocols could not be used to isolate peanut DNA of sufficient quality for PCR amplification or Southern hybridization. We have devised a new protocol that uses DEAE-cellulose purification to isolate peanut DNA useful for downstream applications.

引用

页码：393a / 393h

共 14 条

[11] Rether, B., Delmas, G., Laouedj, A., Isolation of polysaccharide-free DNA from plants (1993) Plant Mol Biol Reptr, 11, pp. 333-337
[12] Rogers, S.O., Bendich, A.J., Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues (1985) Plant Mol Biol, 5, pp. 69-76
[13] Scott, O.R., Bendich, A.J., Extraction of DNA from plant tissues (1988) Plant Mol Biol Manual, A6, pp. 1-10
[14] Sharma, K.K., Anjaiah, V., An efficient method for the production of transgenic plants of peanut (Arachis hypogaea L.) through Agrobacterium tumefaciens-mediated genetic transformation (2000) Plant Sci, 159, pp. 7-19

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