African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12392

Full Length Research Paper

Nuclear isolation and purification using SDS/urea (NIPSU) method for efficient and rapid extraction of high-purity genomic DNAs from Jatropha curcas L: A comparative analysis of DNA isolation protocols

Yoshihiko Nanasato
  • Yoshihiko Nanasato
  • School of Agricultural, Biological and Environmental Sciences, Faculty of Agriculture, Tottori University, 4-101 Koyama-Minami, Tottori 680-8553, Japan.
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Ryo Uenoyama
  • Ryo Uenoyama
  • School of Agricultural, Biological and Environmental Sciences, Faculty of Agriculture, Tottori University, 4-101 Koyama-Minami, Tottori 680-8553, Japan.
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Kaede Tomooka
  • Kaede Tomooka
  • School of Agricultural, Biological and Environmental Sciences, Faculty of Agriculture, Tottori University, 4-101 Koyama-Minami, Tottori 680-8553, Japan.
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Atsushi Kato
  • Atsushi Kato
  • The United Graduate School of Agricultural Sciences, Tottori University, 4-101 Koyama-Minami, Tottori 680-8553, Japan.
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Kinya Akashi
  • Kinya Akashi
  • School of Agricultural, Biological and Environmental Sciences, Faculty of Agriculture, Tottori University, 4-101 Koyama-Minami, Tottori 680-8553, Japan.
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  •  Received: 17 November 2017
  •  Accepted: 20 July 2018
  •  Published: 08 August 2018

Abstract

Jatropha curcas L. (Jatropha) has gained popularity for its potential use in biodiesel production in arid regions, and its yield improvement by molecular breeding has been anticipated. However, Jatropha is known for its recalcitrance in the extraction of pure genomic DNAs. In this study, four DNA extraction methods were comparatively examined for their efficiency and quality of genomic DNA isolations from Jatropha. Consequently, one method, designated as nuclear isolation and purification using SDS/urea (NIPSU) method was found to be the most effective for rapid preparation of high yield and quality genomic DNA from Jatropha leaves. NIPSU method allowed extraction of genomic DNAs from leaves, stems, and roots tissues of Jatropha in high yield (6.8 to 24.5 µg gFW-1), with spectroscopically high purity (A260/A280 and A260/A230 >1.8). NIPSU method was also applicable to the dehydrated leaves for genomic DNA preparations. The isolated genomic DNA was amenable for downstream molecular procedures such as polymerase chain reaction, restriction enzyme cutting and Southern blotting analyses.

 

Key words: Jatropha curcas L., arid region, genomic DNA isolation, nuclear isolation, polymerase chain reaction, Southern blotting analysis.