Extraction of Jute Genomic DNA: Difficulties and Solutions

  • A. K. M. Shahadat Hossain Bangladesh Jute Research Institute
  • Md. Abdul Latif Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202
  • Bipul Kumar Biswas Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202
  • Saidin Saclain Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202
  • Md Salahuddin Department of Physiology, Bangladesh Agricultural University, Mymensingh-2202
  • M. Abu Sayed Department of Biochemistry and Molecular Biology, Hajee Mohammad Danesh Science and Technology University, Dinajpur-500
  • Md. Mahboob Hussain Bangladesh Jute Research Institute
  • Sheik Md Moniruzzaman Department of Biochemistry and Molecular Biology, Bangladesh Agricultural University, Mymensingh-2202
  • Md. Shahidul Islam Bangladesh Jute Research Institute
Keywords: Jute, DNA extraction, phenolic compounds, RAPD

Abstract

The plant parts of jute are composed of highly viscous substances and phenolic compounds which make nuisance to extract good quality genomic DNA (gDNA). Here different methods viz., phenol chloroform isoamylalcohol extraction and ethanol precipitation method; potassium acetate method and Cetyl trimethylammonium bromide (CTAB) method were applied for isolating the jute gDNA from leaves of 14 and 40 day’s plants. All of these methods were unable to extract good quality gDNA from jute leaves. That is why, in this experiment, the concentration and chemical compositions of CTAB method were modified for obtaining good quality jute gDNA. Polyvinylpyrrolidone (PVP) was added in the CTAB extraction buffer and β mercaptoethanol was used while grinding the leaf tissues with CTAB extraction buffer. It was found that good quality gDNA was obtained using modified CTAB method from 14 day’s plant’s leaves but low quality gDNA was obtained from 40 day’s plant’s leaves. These were confirmed by 0.8% agarose gel electrophoresis. Considering the visual quality of the banding patterns and their reproducibly MHR24 (5/-TTCCCTCCCA-3/) and MHR21 (5-/CCCGAAGCGA-3/) primers were selected out of 8 Random Amplified Polymorphic DNA (RAPD) primers for PCR. A total number of 11 loci were identified by these RAPD primers. Popgen32 software was used for analyzing the RAPD data. The number of polymorphic loci was one and the percentage was 9.09 which stated that a low level of genetic variations was existed among the jute accessions. In the dendrogram, jute accession CC875 (C. capsularis) was grouped in one cluster while CC894 (C. capsularis) and CC896 (C. capsularis) accessions were grouped in another cluster.

DOI: http://dx.doi.org/10.3126/ijasbt.v2i4.11320

Int J Appl Sci Biotechnol, Vol. 2(4): 516-520

 

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Abstract
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PDF
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Published
2014-12-25
How to Cite
Hossain, A. K. M. S., Latif, M. A., Biswas, B. K., Saclain, S., Salahuddin, M., Sayed, M. A., Hussain, M. M., Moniruzzaman, S. M., & Islam, M. S. (2014). Extraction of Jute Genomic DNA: Difficulties and Solutions. International Journal of Applied Sciences and Biotechnology, 2(4), 516-520. https://doi.org/10.3126/ijasbt.v2i4.11320
Section
Research Articles