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Molecular Profiling of Dendrocalamus asper and D. Hamiltonii Using RAPD, ISSR and SSR Markers - Effects of DNA Extraction Methods on PCR Amplification
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Extraction of good quality DNA is the most essential requirement for molecular analysis of any species. Five protocols for DNA extraction were compared to assess their effects on amount of DNA recovered, its purity and efficacy in PCR amplification using RAPD, ISSR and SSR markers in two species of Dendrocalamus asper. and D. hamiltonii. Leaf tissues from in vitro cultures, tissue culture raised plants in green house,and young and mature leaves from field grown plants of the two species were subjected to five DNA extraction methods using the protocols. Significant differences in DNA quantity and quality between samples and protocols were observed. Leaves of tissue culture raised plants growing in the green house gave best DNA yield (5681, 5015 ng/ μl)followed by in vitro shoot cultures (4645, 4379 ng/ μl), and young (2720, 1801 ng/ μl) and mature leaves (1800, 1368 ng/ μl) from field grown plants of D. hamiltonii and D. asper respectively.The concentration of DNA obtained was more for D. hamiltonii samples than the corresponding D. asper tissues.
Keywords
Bamboo, Dendrocalamus, DNA Extraction Methods, PCR Efficiency, Quality and Quantity of DNA
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- Alaey, M., Naderi, R., Vezvaei, A., Khalighi, A. and Salami, A. (2005). Comparing study between four different methods of genomic DNA extraction from Cyclamen persicum Mill., Intl. J. Agric. Biol., 7: 882-884.
- Biradar, D.P., Patil, V.C., Kuruvinashetti, M.S., Biradar, M.D., Patil, S.V. and Yaradoni, S.N. (2005). Characterization of bamboo elite clones from Western Ghats of India using RAPD markers; In: The role of biotechnology, Villa Gualino, Turin, Italy, pp. 155-156.
- Bokszczanin, K. and Prazybyla, A.A. (2006). Copper (II) acetate improves the quality of pear (Pyrus) DNA during extraction. Plant Mol. Biol. Rep., 24: 249a-249d.
- Bushra, C., Afshan, Y., Tayyab,H. and Riazuddin, S. (1999). Mini-scale genomic DNA extraction from cotton. Plant Mol. Biol. Rep., 17: 1-7.
- Cankar, K., Stebih, D., Dreo, T., Zel, J. and Gruden, K. (2006). Critical points of DNA quantification by real-time PCR - effects of DNA extraction method and sample matrix on quantification of genetically modified organisms. BMC Biotechnol, 6: 37-51.
- Chiu, W.B., Lin, C.H., Chang, C.J., Hsieh, M.H. and Wang, A.Y. (2006). Molecular characterization and expression of four cDNAs encoding sucrose synthase from green bamboo Bambusa oldhamii. New Phytol, 170: 53-63.
- Csaikl, U.M., Bastian, H., Brettschneider, R., Gauch, S., Meir, A., Schauerte, M., Scholz, F., Sperisen, C., Vornam, B. and Ziegenhagen, B. (1998). Comparative analysis of different DNA extraction protocols: A fast, universal maxi-preparation of high quality plant DNA for genetic evaluation and phylogenetic studies. Plant Mol. Biol. Rep, 16: 69-86.
- Das, M., Bhattacharya, S. and Pal, A. (2005). Generation and characterization of SCARs by cloning and sequencing of RAPD products: A Strategy for Species-specific Marker Development in Bamboo. Ann. Bot, 95: 835-841.
- Dellaporta, S.L., Wood, J. and Hicks, J.B. (1983). A plant DNA minipreparation: version II. Plant Mol. Biol. Rep., 1: 19-21.
- Doyle, J.J. and Doyle, J.L. (1990). Isolation of plant DNA from fresh tissue. Focus, 12: 11-15.
- Fredricks, D.N., Smith, C. and Meier, A. (2005). Comparison of Six DNA Extraction Methods for Recovery of Fungal DNA as Assessed by Quantitative PCR. J. Clinical Microbiol, 43: 5122-5128.
- Ghosh, S., Waikhom, S.D., Mandi, S.S. and Talukdar, N.C. (2011). Amplified Fragment Length Polymorphism based study of phylogenetic relationship and genetic variability among some edible Bamboo species of North-East India. J. Plant Mol. Biol. Biotechnol, 2: 8-15.
- Hunter, I.R. (2003). Bamboo resources, uses and trade: the future? J. Bamboo Rattan 2: 329.
- Jenderek, M.M., Shierenbeck, K.A. and Olney, A.J. (1997). Development of random amplified polymorphic DNA markers characteristic of Hibiscus rosa-sinensis and H. syriacus. Center for Irrigation Technology, California State University, USA.
- Kadkhodaee, S. (2003). A simple method for nucleic acid extraction from plants containing high polysaccharide and polyphenol rd components: Optimizing DNA extraction method in almonds. 3 Congress of Biotechnology, Iran.
- Kalia, R.K., Rai, M.K., Kalia, S., Singh, R. and Dhawan, A.K. (2011). Microsatellite markers: an overview of the recent progress in plants. Euphytica, 177: 309-334.
- Krizman, M., Jakse, J., Baricevic, D., Javornik, B. and Prosek, M. (2006). Robust CTAB-activated charcoal protocol for plant DNA extraction. Acta Agric. Slovenica, 87: 427-433.
- Lin, X.C., Ruan, X.S., Lou, Y., Guo, X.Q. and Fang, W. (2009). Genetic similarity among cultivars of Phyllostachys pubescens. Plant Syst. Evol., 277: 67-73.
- Lodhi, M.A., Ye, G.N., Weeden, N.F. and Reisch, B.I. (1994). A simple and efficient method DNA extraction from grapevine caltivars and vitis species. Plant Mol. Biol. Rep., 12: 6-13.
- Marulanda, M.L., Lopez, A.M. and Claroz, J.L. (2007). Analyzing the genetic diversity of Guadua spp. in Colombia using rice and sugarcane microsatellites. Crop Breed. Appl. Biotech, 7: 43-51.
- McClelland, M. and Welsh, J. (1994). DNA fingerprinting by arbitrarily primed PCR. PCR Methods Applic., 4: s59-s65.
- Murray, M. and Thompson, W.F. (1980). Rapid isolation of molecular weight plant DNA. Nucleic Acid Res., 8: 4321-4325.
- Nayak, S., Rout, G.R. and Das, P. (2003). Evaluation of the genetic variability in bamboo using RAPD markers. Plant Soil Environ., 49: 24-28.
- Nguyen, T.C.T., Son, R., Raha, A.R., Lai, O.M. and Michael, C.W.V.L. (2009). Comparison of DNA extraction efficiencies using various methods for the detection of genetically modified organisms (GMOs). Int. Food Res. J., 16: 21-30.
- Pandey, R.N., Adams, R.P. and Flournoy, L.E. (1996). Inhibition of random amplified polymorphic DNAs (RAPDs) by plant polysaccharides. Plant Mol. Biol. Rep., 14: 17-22.
- Reichardt, M.J. and Rogers, S.J. (1993). Plant DNA isolation using CTAB; In: Current protocols in molecular biology (FM Ausubel, R Brent, RE Kingston, DD Moore, JD Seidman, JA Smith, K Struhl Edts.), New York, USA: John Wiley and Sons, Supplement 22.
- Romman, S.A. (2011). Comparison of methods for isolating high quality DNA from sage (Salvia officinalis). J. Med. Plants Res., 5: 938-941.
- Sambrook, J. and Russel, D.W. (2001). Molecular cloning: a laboratory manual, Third Edition (Cold Spring Harbor Laboratory Press, New York).
- Sarwat, M., Negi, M.S., Lakshmikumaran, M., Tyagi, A.K., Das, S. and Srivastava, P.S. (2006). A standardized protocol for the genomic DNA isolation from Terminalia arjuna for genetic diversity analysis. Elec. J. Biotech., 9: 86-91.
- Talebee, M. (2003). Genetic diversity in pomegranate cultivars in Iran by RAPD marker. 3 Congress of Biotechnology, Iran.
- Yusuf, M. and Muranaka, S. (2009). Comparison of DNA extraction methods for application of marker assisted selection for Striga resistance into national cowpea breeding programmes; Proc. African Crop Sci. Conf., 9: 555-557.
- Ziegenhagen, B., Guillemaut, P. and Scholz, F. (1993). A procedure for mini-preparation of genomic DNA from needles of silver fir (Abies alba Mill.). Plant Mol. Biol. Rep., 11: 117-121.
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