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Authors Gurudeeban, S; Ramanathan, T; Satyavani, K
Author Full Name Gurudeeban, S.; Ramanathan, T.; Satyavani, K.
Title Standardization of Genomic DNA Isolation and Molecular Identification of Rhizophora Sp. using Non-Coding trn L Gene
Source RESEARCH JOURNAL OF BIOTECHNOLOGY
Language English
Document Type Article
Author Keywords Amplification; mangrove; non-coding gene; polyvinylpyrrolidone; Rhizophora
Keywords Plus RAPD
Abstract DNA extraction is difficult in mangrove plants because of phenolic compounds and secondary metabolites that interfere with DNA isolation procedures and subsequent applications such as DNA restriction, amplification and cloning. We developed a simple, rapid and efficient method for isolating genomic DNA from leaves of Rhizophora mucronata and Rhizophora apiculata that is free from polysaccharides and polyphenols. This newly developed protocol includes the use of 2 M NaCl, 0.1 g of polyvinylpyrrolidone (PVP), 3% mercaptoethanol, 1/10 volume of sodium acetate and 70% ethanol in the extraction as well as reducing the centrifugation times during the separation and precipitation of the DNA. Isolated genomic DNA showed high purity and high quantity. The purity of isolated genomic DNA was confirmed by agarose gel electrophoresis. The use of tRNA Leu (trn L) genes also as RFLP markers to precise the genetic diversity and results reveal the degree of polymorphism and to detect similarities among genotypes. The successful double stranded amplification and partial sequences were obtained for the study plants, the size of the trn L (UAA) intron ranging from 587bp for R. mucronata (GenBank Acc.No.: JF318960) and 538bp for R. apiculata (GenBank Acc.No.: JF318959).
Author Address [Gurudeeban, S.; Ramanathan, T.; Satyavani, K.] Annamalai Univ, Fac Marine Sci, Ctr Adv Study Marine Biol, Parangipettai 608502, Tamil Nadu, India
Reprint Address Gurudeeban, S (corresponding author), Annamalai Univ, Fac Marine Sci, Ctr Adv Study Marine Biol, Parangipettai 608502, Tamil Nadu, India.
E-mail Address ramanathanscholars@gmail.com
ResearcherID Number Thiru, Ramanathan/M-2683-2019; Kaliamurthi, Satyavani/D-9417-2012; Selvaraj, Gurudeeban/D-9415-2012
ORCID Number Thiru, Ramanathan/0000-0001-8171-8433; Kaliamurthi, Satyavani/0000-0002-3604-8810; Selvaraj, Gurudeeban/0000-0002-7223-3853
Funding Agency and Grant Number Faculty of Marine Sciences, Annamalai University, Parangipettai, Tamil Nadu, India
Funding Text The authors are gratefully acknowledged to the Director and Dean, Faculty of Marine Sciences, Annamalai University, Parangipettai, Tamil Nadu, India for providing all support during the study period.
Publisher RESEARCH JOURNAL BIOTECHNOLOGY
Publisher City INDORE
Publisher Address SECTOR A-80, SCHEME NO 54, VIJAY NAGAR, A B ROAD, INDORE, 452 010 MP, INDIA
ISSN 2278-4535
29-Character Source Abbreviation RES J BIOTECHNOL
ISO Source Abbreviation Res. J. Biotechnol.
Publication Date MAY
Year Published 2013
Volume 8
Issue 5
Beginning Page 44
Ending Page 47
Page Count 4
Web of Science Category Biotechnology & Applied Microbiology
Subject Category Biotechnology & Applied Microbiology
Document Delivery Number 131BH
Unique Article Identifier WOS:000317965300008
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