Abstract
Cotton is a stubborn plant for genomic DNA isolation due to its high-level of polyphenolics, polysaccharides and secondary metabolites. Genomics and molecular biology studies require high quality and large quantity of DNA. We have standardized an efficient miniprep protocol for cotton genomic DNA isolation, which not only provides higher yield from 800 to 1400 µg of DNA from 200 to 300 mg of fresh leaf tissue but also provide excellent purity. The DNA is amenable to all elementary enzymatic preparations, PCR techniques, Southern blotting and to high end genomic studies. The technique does not require liquid nitrogen, needs small amount of sample, less time, fewer chemicals and one can process up to 100 samples per day. The genomic DNA extracted was good for transgenic event characterization and marker assisted selection.
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References
Abd-Elsalam KA, Asran-Amal A, El-Samawaty AMA (2007) Isolation of high-quality DNA from cotton and its fungal pathogens. J Plant Dis Protect 114(3):113–116
Chaudhary B, Yasmeen A, Husnain T, Riazuddin S (1999) Mini-scale genomic DNA extraction from cotton. Plant Mol Biol Rep 17:1–7
Dabo MS, Mitchell DE, Melcher U (1993) A method for isolation of nuclear DNA from cotton (Gossypium) leaves. Anal Biochem 210:34–38
Horn EC, Kumpatla SP, Patterson KA, Gupta M, Thompson SA (2004) Improved high-throughput sunflower and cotton genomic DNA extraction and PCR fidelity. Plant Mol Biol Rep 22:83a–83i
Kalbande BB, Sutar SR, Sable SV, Meshram MD, Warade J (2012) Principles of DNA extraction and role of various components used in DNA extraction. Agrobios Newsl 12(01):06–08
Li H, Luo J, Hemphill JK, Wang JT, Gould JH (2001) A rapid and high yielding DNA miniprep for cotton (Gossypium spp.). Plant Mol Bio Rep 19:1–5
Paterson AH, Brubaker CL, Wendel JF (1993) A rapid method for extraction of cotton (Gossypium ssp.) genomic DNA suitable for RFLP or PCR analysis. Plant Mol Biol Rep 11:122–127
Permingeat HR, Romagnoli MV, Vallejos RH (1998) A simple method for isolating high yield and quality DNA from cotton (Gossypium hirsutum L) leaves. Plant Mol Biol Rep 16:1–6
Rao PS, Kumar PS, Sonti RV (2013) Excised radicle tips as a source of genomic DNA for PCR-based genotyping and melting curve analysis in cotton. J Biosci 38(1):167–172
Roychowdhury R, Karmakar J, Dey N (2012) PCR-compatible genomic DNA isolation from different tissues of rice (Oryza sativa) for SSR fingerprinting Eurasia. J Biosci 6:85–90
Saha S, Callahan FE, Dollar DA, Creech JB (1997) Effect of lyophilisation of Cotton tissue on quality of extractable DNA, RNA, and protein. J Cotton Sci 1:10–14
Sambrook J, Russell DW (2001) Molecular cloning: a laboratry manual. Cold Spring Harbour Laboratory Press, New York
Xin Z, Chen J (2012) A high throughput DNA extraction method with high yield and quality. Plant Methods 8:26
Zhang J, Stewart JM (2000) Economical and rapid method for extracting cotton genomic DNA. J Cotton Sci 4:193–201
Acknowledgments
The authors wish to acknowledge with thanks to the Head, Department of Biotechnology, S.G.B. Amravati University, Amravati, Maharashtra, 444602, India for providing workbench and his constant encouragement.
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Kalbande, B.B., Patil, A.S. & Chakrabarty, P.K. An efficient, simple and high throughput protocol for cotton genomic DNA isolation. J. Plant Biochem. Biotechnol. 25, 437–441 (2016). https://doi.org/10.1007/s13562-016-0360-z
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DOI: https://doi.org/10.1007/s13562-016-0360-z