Total views : 1465
A Method for Small Scale Genomic DNA Isolation from Chickpea (Cicer arietinum L.) Suitable for Molecular Marker Analysis
We describe a DNA isolation procedure for chickpea (Cicer arietinum L.) which is rapid and less expensive without involving ultra centrifugation or column purification steps. DNA preparation obtained from the present study was essentially suitable for PCR analysis which is one of the key steps in crop improvement programme through marker development and genetic engineering techniques. The yield of DNA ranged from 0.595- 5.550 μg/ml and the purity ratio was between 1.025- 2.010 indicating minimum levels of contaminating metabolites. The present protocol offers as a reliable, and consistent DNA isolation method for chickpea that yields large amount of pure&intact DNA.
DNA Isolation, Cicer arietinum, PCR Analysis
- Do N and Adams RP (1991) A simple technique of removing plant polysaccharides contaminants from DNA. Biotechniques. 10, 162-166.
- Doyle JJ and Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19, 11-15.
- Doyle JJ and Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus. 12, 13-15.
- Fang G, Hammer S and Groumet R (1992) A quick and inexpensive method for removing polysaccharides from plant genomic DNA. Biofeedback. 13, 52-54.
- Lodhi MA, Ye GN and Weeden NF (1994) A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol. Biol. Rep. 12, 6-13.
- Loomis MD (1974) Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Methods Enzymol. 31, 528–544.
- Mace ES, Buhariwalla HK and Crouch JH (2003) A high-throughput DNA extraction protocol for tropical molecular breeding programs. Plant Mol. Biol. Rep. 21, 459a-459h.
- Merlo DJ and Kemp JD (1976) Effect of polysaccharides on kinetics of DNA. Plant Physiol. 58, 1522-1526.
- Pandey RN, Adams RP and Flournoy LE (1996) Inhibitions of random amplified poly-morphic DNAs (RAPDs) by plant polysaccharides. Plant Mol. Biol. Rep. 14, 17-22.
- Paterson AH, Brubaker CL and Wendel JF (1993) A rapid method for extraction of cotton (Gossypium spp.) genomic DNA suitable for RFLP and PCR analysis. Plant Mol. Biol. Rep. 11, 122-127.
- Pikart MJ and Villeponteau B (1993) Suppression of PCR amplification by high levels of RNA. Biotech.14, 24-25.
- Porebski S, Bailey LG and Baum BR (1997) Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol. Biol. Rep. 15, 8–15.
- Sambrook J, Fritch EF and Maniatis T (1989) Molecular cloning: A laboratory manual. Cold Springer Harbor Laboratory Press, Cold Spring Harbor, NY.
- Scott KD and Playford J (1996) DNA extraction technique for PCR in rain forest plant species. Biotech. 20, 974-978.
- Sharma AD, Gill PK and Singh P (2002) DNA isolation from dry and fresh samples of polysaccharide rich plants. Plant Mol. Biol. Rep. 20, 415a– 415f.
- Sharma KK, Lavanya M and Anjaiah V (2000) A method for isolation and purification of peanut genomic DNA suitable for analytical applications. Plant Mol. Biol. Rep. 18, 393a-393h.
- Varshney RK, Horres R, Molina C, Nayak S, Jungmann R, Swamy P, Winter P, Jayashree B, Kahl G and Hoisington DA (2007) Extending the repertoire of microsatellite markers for genetic linkage mapping and germplasm screening in chickpea. J. SAT Agri. Res. 5(1), 1-3.
- Weishing K, Nybom H, Wolff K and Meyer W (1995) DNA isolation and purification. In: DNA fingerprinting in plants and fungi, CRC Press, Boca Raton, Florida, pp: 44-59.
- There are currently no refbacks.
This work is licensed under a Creative Commons Attribution 3.0 License.