An Efficient Method for High Quality RNA Extraction from Moringa oleifera
Journal of Plant Sciences
Volume 5, Issue 2, April 2017, Pages: 68-74
Received: Feb. 19, 2017; Accepted: Feb. 28, 2017; Published: Mar. 28, 2017
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Authors
Preethi Praba Umesh, Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India
Mohd Akram Ansari, Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India; Division of Genetics and Plant Molecular Biology, CSIR-NBRI, Rana Pratap Marg, Lucknow, Uttar Pradesh, India
Ganapathi Sridevi, Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India
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Abstract
Moringa oleifera, the Miracle Tree is rich in all nutrients and minerals. It has prominent distribution of secondary metabolites like polysaccharides, phenols and mucilage, which makes extraction of RNA quite difficult. A high quality and pure RNA is prerequisite for the study of high through put transcriptomics and functional genomics. A protocol for isolation of highly qualitative and quantitative RNA from M. oleifera was optimized by comparing five different methods like Trizol, Guanidine hydrochloride, combined Trizol and Guanidine hydrochloride, modified CTAB and hot phenol method. The combined Guanidine hydrochloride and Trizol method gave good yield and pure RNA based on the absorbance A260 value 2.0. The model plant Nicotiana tabacum served as a positive control in which the Trizol method yielded a good quality and quantity RNA. The present study is a preliminary step for studying the function and expression pattern of the genes. This is the first report on the comparison of different RNA extraction methods in M. oleifera to our knowledge.
Keywords
Moringa oleifera, Nicotiana tabacum, RNA Isolation, Trizol, Guanidine Hydrochloride, RT-PCR
To cite this article
Preethi Praba Umesh, Mohd Akram Ansari, Ganapathi Sridevi, An Efficient Method for High Quality RNA Extraction from Moringa oleifera, Journal of Plant Sciences. Vol. 5, No. 2, 2017, pp. 68-74. doi: 10.11648/j.jps.20170502.14
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Copyright © 2017 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
References
[1]
Box MS, Coustham V, Dean C, Mylne JS. 2011. Protocol: A simple phenol based for 96- well extraction of high quality RNA from Arabidopsis. Plant Method 7:7.
[2]
Chan KL, Ho CL, Namasivayam P, Napis S. 2007. A simple and rapid method for RNA isolation from plant tissues with high phenolic compounds and polysaccharides. Protoc Exch 184.
[3]
De Vries S, Hoge H, Bisseling T. 1993. in Plant Molecular Biology manual, edited by SB Gelvin, RA Schiport and DPS Verma. Kluwer Academic Publisher, Dordretch 1-13.
[4]
Forestry/ Fuelwood Research and Development Project (F/FRDP). 1992. Growing multipurpose trees on small farms. Bangkok. Thailand. Winrock international. (http://agrss.sleman.hawaii.edu/onfarm/tree/tree0012.html).
[5]
Fuglie JL. 1999. Natured nutrition for Tropics. Training Manual Church world service, Dakar The multiple attributes of Moringa 172.
[6]
Fuglie JL. 2001. The Miracle tree: Moringa oleifera: Natured nutrition for Tropics. Training Manual Church world service, Dakar, Senegal.
[7]
Gopalan C, Rama sastri BV, Balasubramanian SC. 1971. Nutritive value of Indian foods.
[8]
Hartwell JL. 1967-1971. Plants used against cancer. A Survey. L. Loydia, 30034.
[9]
http://WWW.Moringa.rubiconhealth.Org.
[10]
Jones S, Iannetta M, Woodhead M, Davies V, McNicol J, Taylor A. 1997. The isolation of RNA from raspberry (Rubus idaeus) fruit. Mol Biotechnol 8:219-221.
[11]
Li B, Wang B, Tang K, Wei J. 2006. A simple and convenient approach for isolating RNA from highly viscous plant tissue rich in polysacharrides. Colloid Surf B: Biointerfaces 49:101-5.
[12]
Logemann J, Schell J, Willmitzer L. 1987. Improved method for the isolation of RNA from plant tissues. Ann Biochem 163:16-20.
[13]
Loomis WD. 1974. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Method Enzymol 31:528-544.
[14]
Manuel GC, Francisco MC. 1999. RNA isolation from plant tissues: a practical experience for biological undergraduates. Biochem Educ 27:110-113.
[15]
Pawlowski K, Kunze R, Vries S De, Bisseling T. 1994. Isolation of total, poly(A) and polysomal RNA from plant tissues in S. B. Gelvin and R. A. Schilperoort (ed.) Plant Molecular Biology Manual, Kluwer Academic Publishers, Dordretch.
[16]
Porbeski S, Bailey LG, Baum BR. 1997. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol component. Plant Mol Biol Rep 15:8-25.
[17]
Sambrook J, EF Fritsch, T Maniatis 1989. Molecular cloning: A laboratory manual. Cold spring harbor Laboratory Press Vol 1 and 3.
[18]
Soni R, Murray J AH. 1994. Isolation of intact DNA and RNA from plant tissues. Ann Biochem 218:474.
[19]
Pirro S, Minutolo A, Galgani A, Potesta M, Colizzi V, Montesano C. 2016. Bioinformatics prediction and experimental validation of micromas involved in cross-kingdom interaction. J comput biol 23:1-14.
[20]
Tian Y, Zeng Y, Zhang J, Yang C, Yan L, Wang X, Shi C, Xei J, Dai T, Peng L, Zeng Y, Huan, Xu A, Huang Y, Zhang J, Max, Dong Y, Hao S, Sheng J. 2015. High quality reference genome of drumstick tree (Moringa oleifera), A potential perennial crop. Sci CHINA LIFE Sci 58:627-638.
[21]
Verma SC, Banerji R, Misra G, Nigam SK. 1976. Nutritional value of Moringa. Curr Sci 45:769-770.
[22]
Wang CS, Vodkin L. 1994. Extraction of RNA from tissues containing high levels of procyanidins that bind RNA. Plant Mol Biol Rep 12:132-145.
[23]
Yang G, Zhou R, Tang T, Shi S. 2008. Simple and efficient isolation of high quality total RNA from Hibiscus tiliaceus, a mangrove associate and its relatives. Prep Biochem Biotechnol 38:25-264.
[24]
Zeng Y, Yang T. 2002. RNA Isolation from highly viscous samples rich in polyphenols and polysaccharides. Plant Mol Biol Rep 20:417a–417e.
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