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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 6 (december 2019) : 749-752,   Doi: 10.18805/IJARe.A-5212

Comparative investigation of DNA extraction methods in black gram (Vigna mungo (L.) 

M
M. Prakash
B
B. Priyadharshini
M
M. Vignesh
R
R. Anandan
1Department of Genetics and Plant Breeding, Faculty of Agriculture, Annamalai University, Annamalai Nagar-608 002, Tamil nadu, India.
Cite article:- Prakash M., Priyadharshini B., Vignesh M., Anandan R. (2019). Comparative investigation of DNA extraction methods in black gram (Vigna mungo (L.). Indian Journal of Agricultural Research. 53(6): 749-752. doi: 10.18805/IJARe.A-5212.

ABSTRACT

Isolation of intact, double stranded, pure and non- contaminated genomic DNA is prerequisite for large scale genotyping analysis including DNA-banks. Three methods of DNA isolation (Dellaporta, CTAB and Hi-PurAg DNA isolation kits) from 25 black gram genotypes were compared in terms of the yield, purity, integrity, and stability of extracted DNA. Purity and quantification of isolated DNA samples was confirmed by using the UV nano-spectrophotometer at OD260/280 and the same is confirmed based by agarose gel electrophoresis. The CTAB method showed the best results followed by Hi-PurAg and Dellaporta method. The CTAB DNA extraction method was found to be the most efficient DNA extraction method, capable of providing high quality, pure and stable DNA and could be used for various molecular related works. All the 25 black gram genotypes for this research gave good yield of DNA from the established modified CTAB protocol. 

REFERENCES

  1. Abdel-Latif, A. and Osman, G. (2017). Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize. Plant Methods, 13(1): 1.
  2. Braid, M., Daniels, L. and Kitts, C. (2003). Removal of PCR inhibitors from soil DNA by chemical flocculation. J Microbiol Methods, 52: 389-93.
  3. Da,s D.K., Bhagat, M. and Shree, S. (2016).Agrobacterium mediated transformation of Vigna mungo L. Hepper with Cry1Ac gene for insect resistance. American J. of Plant Sci., 7: 316-325.
  4. Delic, D., Stajkovic, O., Kuzmanovic, D., Rasulic, N., Knezevic, N. and Milicic, B. (2009). The effects of rhizobial inoculation on growth and yield of Vigna mungo L. in Serbian soils. Biotechnology in Animal Husbandry, 25(5-6): 1197-1202.
  5. Dellaporta, S,L., Wood, J. and Hicks, J.B. (1983). A Plant DNA Minipreparation: Version II. Plant Molecular Biology Reporter, 1: 19-21.
  6. Doyle, J.J. and Doyle, J.L. (1990). Isolation of plant DNA from fresh tissue. Focus, 12:13-15.
  7. Kasem, S., Rice, N. and Henry, R. (2008). DNA extraction from plant tissue. In: Plant genotyping II: SNP technology [Henry RJ, editor], CAB International, Wallingford, 219-71.
  8. Kotchoni, S.O., Gachomo, E. W. and Jimenez-Lopez, J. C. (2011). A plant cocktail amenable for PCR-based genetic analysis in Arabidopsis thaliana. Mol. Biol. Rep., 38: 5281-5284
  9. Margam, V.M., Gachomo, E. W., Shukle, J. H. Ariyo, O. O., Manfredo, J., Seufferheld, M. J. and Kotchoni, S. O. (2010). A simplified arthropod genomic-DNA extraction protocol for polymerase chain reaction (PCR)-based specimen identification through barcoding. Mol. Biol. Rep., 37: 3631–3635.
  10. Osman, G.H., Assem, S. K., Alreedy, R. M., El-Ghareeb, D. K., Basry, M. A., Rastogi, A. and Kalaji, H. M. (2015). Development of insect resistance maize plants expressing the chitinase gene of Spodoptera littoralis. Nat Sci Rep., 5:18067-77.
  11. Pervaiz, Z., Turi, N., Khaliq, I., Rabbani, M. and Malik, S. (2011). A modified method for high-quality DNA extraction for molecular analysis in cereal plants. Genet Mol Res J., 10:1669-73.
  12. Porebski, S., Bailey, L. G. and Baum, B. R. (1997). Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol.Biol. Report., 15: 8-15.
  13. Sarwat, M., Negi, M. Lakshmikumaran, M. and Tyagi, M.A. (2006). A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis. Electron J Biotechnol., 9:86-91.
  14. Tharanathan, R.N. and Mahadevamma S. (2003). Grain legumes—a boon to human nutrition. Trends Food Sci Technol, 14: 507-518. 
  15. Verdcourt, B. (1966). The arid corridor between the northeast and southwest areas of Africa. Palaeoecol. Afr., 4:140-144.
  16. Wang, T.Y., Wang, L., Zhang, J. H. and Dong, W. H. (2011). A simplified universal genomic DNA extraction protocol suitable for PCR. Genet. Mol. Res., 10: 519-525. 
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Indian Journal of Agricultural Research
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    Research Article

    volume 53 issue 6 (december 2019) : 749-752,   Doi: 10.18805/IJARe.A-5212

    Comparative investigation of DNA extraction methods in black gram (Vigna mungo (L.) 

    M
    M. Prakash
    B
    B. Priyadharshini
    M
    M. Vignesh
    R
    R. Anandan
    1Department of Genetics and Plant Breeding, Faculty of Agriculture, Annamalai University, Annamalai Nagar-608 002, Tamil nadu, India.
    Cite article:- Prakash M., Priyadharshini B., Vignesh M., Anandan R. (2019). Comparative investigation of DNA extraction methods in black gram (Vigna mungo (L.). Indian Journal of Agricultural Research. 53(6): 749-752. doi: 10.18805/IJARe.A-5212.

    ABSTRACT

    Isolation of intact, double stranded, pure and non- contaminated genomic DNA is prerequisite for large scale genotyping analysis including DNA-banks. Three methods of DNA isolation (Dellaporta, CTAB and Hi-PurAg DNA isolation kits) from 25 black gram genotypes were compared in terms of the yield, purity, integrity, and stability of extracted DNA. Purity and quantification of isolated DNA samples was confirmed by using the UV nano-spectrophotometer at OD260/280 and the same is confirmed based by agarose gel electrophoresis. The CTAB method showed the best results followed by Hi-PurAg and Dellaporta method. The CTAB DNA extraction method was found to be the most efficient DNA extraction method, capable of providing high quality, pure and stable DNA and could be used for various molecular related works. All the 25 black gram genotypes for this research gave good yield of DNA from the established modified CTAB protocol. 

    REFERENCES

    1. Abdel-Latif, A. and Osman, G. (2017). Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize. Plant Methods, 13(1): 1.
    2. Braid, M., Daniels, L. and Kitts, C. (2003). Removal of PCR inhibitors from soil DNA by chemical flocculation. J Microbiol Methods, 52: 389-93.
    3. Da,s D.K., Bhagat, M. and Shree, S. (2016).Agrobacterium mediated transformation of Vigna mungo L. Hepper with Cry1Ac gene for insect resistance. American J. of Plant Sci., 7: 316-325.
    4. Delic, D., Stajkovic, O., Kuzmanovic, D., Rasulic, N., Knezevic, N. and Milicic, B. (2009). The effects of rhizobial inoculation on growth and yield of Vigna mungo L. in Serbian soils. Biotechnology in Animal Husbandry, 25(5-6): 1197-1202.
    5. Dellaporta, S,L., Wood, J. and Hicks, J.B. (1983). A Plant DNA Minipreparation: Version II. Plant Molecular Biology Reporter, 1: 19-21.
    6. Doyle, J.J. and Doyle, J.L. (1990). Isolation of plant DNA from fresh tissue. Focus, 12:13-15.
    7. Kasem, S., Rice, N. and Henry, R. (2008). DNA extraction from plant tissue. In: Plant genotyping II: SNP technology [Henry RJ, editor], CAB International, Wallingford, 219-71.
    8. Kotchoni, S.O., Gachomo, E. W. and Jimenez-Lopez, J. C. (2011). A plant cocktail amenable for PCR-based genetic analysis in Arabidopsis thaliana. Mol. Biol. Rep., 38: 5281-5284
    9. Margam, V.M., Gachomo, E. W., Shukle, J. H. Ariyo, O. O., Manfredo, J., Seufferheld, M. J. and Kotchoni, S. O. (2010). A simplified arthropod genomic-DNA extraction protocol for polymerase chain reaction (PCR)-based specimen identification through barcoding. Mol. Biol. Rep., 37: 3631–3635.
    10. Osman, G.H., Assem, S. K., Alreedy, R. M., El-Ghareeb, D. K., Basry, M. A., Rastogi, A. and Kalaji, H. M. (2015). Development of insect resistance maize plants expressing the chitinase gene of Spodoptera littoralis. Nat Sci Rep., 5:18067-77.
    11. Pervaiz, Z., Turi, N., Khaliq, I., Rabbani, M. and Malik, S. (2011). A modified method for high-quality DNA extraction for molecular analysis in cereal plants. Genet Mol Res J., 10:1669-73.
    12. Porebski, S., Bailey, L. G. and Baum, B. R. (1997). Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol.Biol. Report., 15: 8-15.
    13. Sarwat, M., Negi, M. Lakshmikumaran, M. and Tyagi, M.A. (2006). A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis. Electron J Biotechnol., 9:86-91.
    14. Tharanathan, R.N. and Mahadevamma S. (2003). Grain legumes—a boon to human nutrition. Trends Food Sci Technol, 14: 507-518. 
    15. Verdcourt, B. (1966). The arid corridor between the northeast and southwest areas of Africa. Palaeoecol. Afr., 4:140-144.
    16. Wang, T.Y., Wang, L., Zhang, J. H. and Dong, W. H. (2011). A simplified universal genomic DNA extraction protocol suitable for PCR. Genet. Mol. Res., 10: 519-525. 
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