Evaluation of economical and rapid method of plant DNA extraction for PCR analysis of different crops
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Abstract
In the recent genomic era, polymerase chain reaction (PCR) has become a basic tool in molecular studies and the success of PCR depends upon the template DNA. PCR technique is quite robust and often unnecessary to extract high quality of DNA and hence crude DNA can be used as template for amplification. Therefore, we have evaluated NaOH-Tris DNA extraction method for PCR analysis because this is very simple, time saving and safe without the need to use expensive or rare materials and laboratory apparatus. This method only requires a small amount of leaf tissue, NaOH, Tris, micro tube and plastic pestle. The amplified PCR products showed clear, sharp and uniform bands gave similar results as compared with the modified CTAB method. The DNA obtained is crude contains impurities like protein, RNA but these impurities did not affect PCR amplification. This DNA extraction method is evaluated for brinjal (Solanummelongena L.), chilli (Capsicum annuum L.), rice (Oryza sativa L.) and tomato (Solanumlycopersicum L.) crop. Many other crop plants could also be amplified using the same DNA extraction method for molecular analysis of large samples. Thus, the use of NaOH-Tris method will allow researchers to obtain DNA from plant quickly for use in molecular studies.
Article Details
Article Details
DNA extraction, NaOH, PCR, Tris
Berthoumieu, P. and Meyer, C. (1991). Direct amplification of plant genomic DNA from leaf and root pieces using PCR. Plant Molecular Biology Report, 17: 555-557
Cheung, W. Y., Hubert, N. and Landry, B. S. (1993). A simple and rapid DNA microextraction method for plant, animal, and insect suitable for RAPD and other PCR analyses. PCR Methods and Applications, 3: 69-70
Chunwongse, J., Martin, G. B. and Tanksley, S. D. (1993). Pre-germination genotypic screening using PCR amplification of half-seeds. Theoretical and Applied Genetics, 86: 694-698
Collard, B. C. Y., Das, A., Virk, P. S. and Mackill, D. J. (2007). Evaluation of ‘quick and dirty’ DNA extraction methods for marker-assisted selection in rice (Oryza sativa L.). Plant Breeding, 126: 47-50
Dellaporta, S. L., Wood, J. and Hicks, J. B. (1983). A plant DNA minipreparation: version II. Plant Molecular Biology Report, 1: 19-21
Doyle, J. J. and Doyle, J. L. (1990). Isolation of plant DNA from fresh tissue, Focus, 12: 13-15
Edwards, K., Johnstone, C. and Thompson, C. (1991). A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Research, 19(6): 1349
Guidet, F. (1994). A powerful new technique to quickly prepare hundreds of plant extracts for PCR and RAPD analyses. Nucleic Acids Research, 22: 1772-1773
Haymes, K. M. (1996). Mini-prep method suitable for a plant breeding program. Plant Molecular Biology Report, 14: 280-284
Kwok, S. and Higuchi, R. (1989). Avoiding false positives with PCR. Nature, 339: 237-238
Lange, D. A., Penuela, S., Denny, R. L., Mudge, J., Concibo, V. C., Orf, J. H. and Young, N. D. (1998). A plant DNA isolation protocol suitable for polymerase chain reaction based marker-assisted breeding. Crop Science, 38:217-220
Langridge, U., Schwall, M. and Langridge, P. (1991). Squashes of plant tissue as substrate for PCR. Nucleic Acids Research, 19: 6954-6956
Lassner, M. W., Peterson, P. and Yoder, J. I. (1989). Simultaneous amplification of multiple DNA fragments by polymerase chain reaction in the analysis of transgenic plants and their progeny. Plant Molecular Biology
Reporter, 7: 116-128
Minamiyama, Y., Tsuro, M. and Hirai, M. (2006). An SSR-based linkage map of Capsicum annuum. Molecular Breeding, 18: 157-169
Nunome, T., Negoro, S., Kono, I., Kanamori, H., Miyatake, K., Yamaguchi, H., Ohyama, A. and Fukuoka, H. (2009). Development of SSR markers derived from SSR-enriched genomic library of eggplant (Solanum melongena L.). Theoretical and Applied Genetics, 119: 1143-1153
Oard, J. H. and Dronavalli, S. (1992). Rapid isolation of rice and maize DNA for analysis by random-primer PCR. Plant Molecular Biology Report, 10: 236-241
Saghai-Maroof, M. A., Soliman, K. M., Jorgensen, R. A. and Allard, R. W. (1984). Ribosomal DNA spacer-length polymorphism in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Sciences, USA, 81: 8014-8019
Septiningsih, E. M., Pamplona, A. M., Sanchez, D. L., Neeraja, C. N., Vergara, G. V., Heuer, S., Ismail, A. M. and Mackill, D. J. (2009). Development of submergence tolerant rice cultivars: the Sub1 locus and beyond. Annals of Botany, 103: 151-160
Thomson, D. and Henry, R. (1995). Single-step protocol for preparation of plant tissues for analysis by PCR. Biotechniques, 19: 394-400
Wang, H., Qi, M. and Cutler, A. J. (1993). A simple method of preparing plant samples for PCR. Nucleic Acids
Research, 21: 4153-4154.
Warner, P., Karakousis, A., Schiemann, A., Eglinton, J., Langridge, P. and Barr, A. (2001). An investigation of a rapid DNA extraction method for routine MAS in the S.A. Barley Improvement Program. Proceeding of the 10th Australian Barley Tech Symposium
Zhang, J. and Stewart, J. M. (2000). Economical and rapid method for extracting cotton genomic DNA. The
Journal of Cotton Science, 4: 193-201
Zhang, C., Liu, L., Wang, X., Vossen, J., Li, G., Li, T., Zheng, Z., Gao J., Guo, Y., Visser, R. J. F., Li, J., Bai, Y. and Du, Y. (2014). The Ph-3 gene from Solanum pimpinellifolium encodes CC NBS LRR protein conferring resistance to Phytophthora infestans. Theoretical and Applied Genetics, 127: 1353-1364
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