Development of a Competent and Trouble Free DNA Isolation Protocol for Downstream Genetic Analyses in Glycine Species

Extraction of deoxyribose nucleic acid (DNA) from plants is preliminary step in molecular biology. Fast and cost effective genomic DNA isolation from Glycine species for downstream application is a major bottleneck. Here we report a high throughput and trouble free method for genomic DNA extraction from leaf and seeds of Glycine species with high quality and quantity. Protocol reports the optimization by employing different concentrations of CTAB and PVP in extraction buffer. Efficiency of optimized protocol was compared with frequently used DNA extraction methods. Wide adoptability and utility of this protocol was confirmed by DNA extraction from leaves as well as seeds of G. max, G. soja, G. tomentella and G. latifolia. Extracted DNA was successfully subjected to PCR amplification of five microsatellite markers and four putative glycosyltransferase genes. DNA extraction protocol is reproducible, trouble free, rapid and can be adopted for plant molecular biology applications.

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Bhau BS, Gogoi G, Baruah D, et al. 2015. Development of an effective and efficient DNA isolation method for Cinnamomum species. Food Chemistry, 188: 264-270.

Bressan EA, Rossi ML, Gerald LT, et al. 2014. Extraction of high- quality DNA from ethanol-preserved tropical plant tissues. BMC Research Notes, 7: 268.

Chang S, Thurber CS, Brown PJ, et al. 2014. Comparative mapping of the wild perennial Glycine latifolia and soybean (G. max) reveals extensive chromosome rearrangements in the genus Glycine. PLoS ONE, 9: e99427.

Demeke T, Adams RP. 1992. The effects of plant polysaccharides and buffer additives on PCR. Biotechniques, 12: 332-334.

Dolatabadi JEN, Kashanian S. 2010. A review on DNA interaction with synthetic phenolic food additives. Food Res. Int., 43: 1223-1230.

Doyle JJ, Doyle JI. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull., 19: 11- 15.

Drabkova L, Kirschner JAN, Vlcek C. 2002. Comparison of seven DNA extraction and amplification protocols in historical herbarium specimens of Juncaceae. Plant Mol. Biol. Reports, 20: 161-175.

Fang G, Hammar S, Grumet R. 1992. A quick and inexpensive method for removing polysaccharides from plant genomic DNA. Biotechniques, 13: 52-54.

Guo J, Wang Y, Song C, et al. 2012. A single origin and moderate bottleneck during domestication of soybean (Glycine max): implications sequences. Ann. Bot., 106: 505-514. and nucleotide

Haberer G, Fischer TC, Torres-Ruiz RA. 1996. Mapping of the nucleolus organizer region on chromosome 4 in Arabidopsis thaliana. Mol. Genet. Genomics, 250: 123-128.

Khanuja SP, Shasany AK, Darokar MP, et al. 1999. Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Mol. Biol. Reports, 17: 74-74.

Kit YS, Chandran S. 2013. A simple, rapid and efficient method of isolating DNA from Chokanan mango (Mangifera indica L.). Afric. J. Biotech., 9: 5805-5808.

Malencic D, Cvejic J, Miladinovic J. 2012. Polyphenol content and antioxidant properties of colored soybean seeds from Central Europe. Journal of Medicinal Food, 15:89-95.

Maliyakal JE. 1992. An efficient method for isolation of RNA and DNA from plants containing polyphenolics. Nucleic Acids Res., 20: 2381.

Manoj K, Tushar B, Sushama C. 2007. Isolation and purification of genomic DNA from Black Plum (Eugenia jambolana Lam.) for analytical applications. Int. J. Biotech. & Biochem., 3: 49-55.

Mesquita ACO, Figueiro AA, Couto KR, et al. 2015. Optimization of soybean DNA extraction under different storage and development periods. J. Biosci., 31: 1102-1106.

Moreira PA, Oliveira DA. 2011. Leaf age affects the quality of DNA extracted from Dimorphandra mollis (Fabaceae), a tropical tree species from the Cerrado region of Brazil. Genet. & Mol. Res., 10: 353-358.

Pirttila AM, Hirsikorpi M, Kamarainen T, et al. 2001. DNA isolation methods for medicinal and aromatic plants. Plant Mol. Bio. Rep., 19: 273-273.

Porebski S, Bailey LG, Baum BR. 1997. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol. Bio. Rep., 15: 8-15.

Sahu SK, Thangaraj M, Kathiresan K. 2012. DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. Mol. Bio., 1-6.

Sharma AD, Gill PK, Singh P. 2002. DNA isolation from dry and fresh samples of polysaccharide-rich plants. Plant Mol. Bio. Rep., 20: 415-415.

Suman PSK, Ajit KS, Darokar MP, Kumar S. 1999. Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Mol. Bio. Rep. 17: 1-7.

Watson, RR. 2014. Polyphenols in Plants: Isolation, Purification and Extract Preparation. London: Academic Press.
Türk Tarım - Gıda Bilim ve Teknoloji dergisi-Cover
  • ISSN: 2148-127X
  • Yayın Aralığı: Aylık
  • Başlangıç: 2013
  • Yayıncı: Turkish Science and Technology Publishing (TURSTEP)
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