International network for natural sciences – research journal
  • mendeley icon
  • linkedin icon
  • google plus icon
  • twitter icon
  • google scholar icon
  • facebook icon

Extraction of purified plants gDNA free of Secondary metabolites

By: Shahid Ali Khan, Niaz Ali, Gandeka Mamadou, Qinzheng Liu, Ruirong Zhuang, Hua Chen

Key Words: Shahid Ali Khan*, Niaz Ali, Gandeka Mamadou, Qinzheng Liu, Ruirong Zhuang, Hua Chen

Int. J. Biosci. 10(3), 223-231, March 2017.

DOI: http://dx.doi.org/10.12692/ijb/10.3.223-231

Abstract

Extraction of high-quality gDNA from plants is a basic step in molecular biology research. In some plant species like peanut getting the genomic DNA in its purified form is a very challenging job as compared to other crops. The reason behind which is the coexistence of different secondary metabolite impurities i.e., polyphenol and polysaccharides with genomic DNA. In this paper, we have optimized a protocol for extracting high-quality gDNA which is amenable to various improvement studies like PCR amplification and other molecular research. The OD 260/280 ratio for the extracted DNA was found within the range of 1.8 to 1.99 while OD 260/230 ratio was between 2.0 to 2.20, using Nano Drop ND-2000/2000C Spectrophotometer. Similarly, the concentration of the extracted DNA for all the extracted samples was greater than 300 ng/ul. The obtained results through our current method indicate the fitness of the extracted DNA for using it in PCR amplification and other molecular research. The novelty in our protocol is the use of Buffer A before CTAB treatment of the samples.

| Views 76 |

Extraction of purified plants gDNA free of Secondary metabolites

Cavallari MM, Siqueira MVBM, Val TM, Pavanelli JC, Monteiro M, Grando C, Pinheiro JB, Zucchi MI, Gimenes MA. 2014. A modified acidic approach for DNA extraction from plant species containing high levels of secondary metabolites. Genetic and  Molecular Research3, 6497-6502.

http://dx.doi.org/10.4238/2014

Claudia S, Doris P, Patricio AJ. 1998. Isolation of pinusradiate genomic DNA suitable for RAPD analysis. Plant Molecular Biology Report 16, 1- 8.

http://dx.doi.org/10.1023/A:1007540901981

Dipankar C, Anindya SG, Sampa D. 2006. Small and  large scale genomic DNA  isolation protocol  for  Chickpea  (Cicer arietinum  L.) suitable for  the molecular marker and  transgenic analysis.  African Journal of  Biotechnology 8, 585-589.

Favero AP, Simpson CE, Valls JF, Vello NA. 2006. Study of the evolution of cultivated peanut through cross ability studies among Arachis ipaensis, A. duranensis, and A. hypogaea. Crop Sciences 46, 1546-1552.

http://dx.doi.org/10.2135/cropsci2005.09-0331

Kasem S, Rice N, Henry R. 2008. DNA extraction from plant tissue. In: Henry RJ, editor. Plant genotyping II: SNP technology. Wallingford: CAB International; 2008. 219-71 P.

http://dx.doi.org/10.1079/9781845933821.0000

Kotchoni SO, Gachomo EW. 2009. A rapid and hazardous reagent free protocol for genomic DNA extraction suitable for genetic studies in plants. Molecular  Biology Reporter 36, 1633-1636.

http://dx.doi.org/10.1007/s11033-008-9362-9

Kotchoni SO, Gachomo EW, Jimenez-Lopez JC. 2011. A plant cocktail amenable for PCR-based genetic analysis in Arabidopsis thaliana. Molecular  Biology Reporter 38, 5281-5284.

http://dx.doi.org/10.1007/s11033-011-0677-6

Li YX, Su ZX, Chen F. 2002. Rapid extraction of genomic DNA from leaves and bracts of dove tree (Davidia involucrata). Plant Molecular Biology Report 20, 185.

http://dx.doi.org/10.1007/BF02799433

Milla SR, Isleib TG, Stalker HT. 2005. Taxonomic relationships among Arachis sect. Arachis species as revealed by AFLP markers. Genome 48, 1-11.

http://dx.doi.org/10.1139/g04-089

Murray MG, Thompson WF. 1980.Rapid isolation of high molecular weight DNA. Nucleic Acids Research 8, 4321- 4325.

Padmesh P, Reji JV, Jinish DM, Seeni S. 2006.Estimation of genetic diversity in varieties of Mucuna pruriens using RAPD. Biology Plantarum 50, 367-372.

Paterson AH, Brubaker CL, Weendel JF. 1993. A rapid method for extraction of Cotton (Gossipium spp) genomic DNA suitable for RFLP or PCR analysis. Plant Molecular Biology Report 11, 122-127.

http://dx.doi.org/10.1007/BF02670470

Robert T, Damayantu M, Jitendra GS. 2003. Asimple and rapid method for isolation of DNA from imbibed embryos of Parkiatimoriana for PCR analysis. Food Agriculture Environment 3, 36-38.

Seijo JG, Lavia GI, Fernandez A, Krapovickas A, Ducasse DA, Moscone EA. 2004. Physical mapping of the 5S and 18S–25S rRNA genes by FISH as evidence that Arachis duranensis and A. ipaensis are the wild diploid progenitors of A. hypogaea (Leguminosae). American Journal of  Botany 91, 1294-1303.

http://dx.doi.org/10.3732/ajb.91.9.1294.

Seijo JG, Lavia GI, Fernandez A, Krapovickas A, Ducasse DA, Bertioli DJ, Moscone EA. .2007. Genomic relationships between the cultivated peanut (Arachishypogaea, Leguminosae) and its close relatives revealed by double GISH. American Journal of  Botany 94, 1963-1971.

http://dx.doi.org/10.3732/ajb.94.12.1963.

Sharma KK, Lavanya M, Anjaia V. 2000. A Method for Isolation and Purification of Peanut Genomic DNA Suitable for Analytical Applications. Plant Molecular Biology Report 18, 393-393.

http://dx.doi.org/10.1007/BF02825068

Shepherd M, Cross M, Stokoe RL, Scott LJ, Jones ME. 2002. High-throughput DNA extraction from forest trees. Plant Molecular Biology Report 20, 425.

http://dx.doi.org/10.1007/BF02772134

Suman PSK, Ajit KS, Darokar MP, Kumar  S. 1999.Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils.Plant Molecular Biology Report 17, 1-7.

http://dx.doi.org/10.1023/A:1007528101452

Shahid Ali Khan, Niaz Ali, Gandeka Mamadou, Qinzheng Liu, Ruirong Zhuang, Hua Chen.
Extraction of purified plants gDNA free of Secondary metabolites.
Int. J. Biosci. 10(3), 223-231, March 2017.
http://www.innspub.net/ijb/extraction-purified-plants-gdna-free-secondary-metabolites/
Copyright © 2017
By Authors and International Network for
Natural Sciences (INNSPUB)
http://innspub.net
brand
innspub logo
english language editing
  • CALL FOR PAPERS
    CALL FOR PAPERS
    Publish Your Article
  • CALL FOR PAPERS
    CALL FOR PAPERS
    Submit Your Article
INNSPUB on FB
Email Update