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:: Volume 7, Issue 27 (9-2017) ::
NCMBJ 2017, 7(27): 55-62 Back to browse issues page
Comparative study of different protocols for isolation of RNA from apple leaves (Malus domestica)
Shahram Montazeri , Shahab Sadat
Department of Plant Breeding, Khuzestan Science and research Branch, Islamic Azad University, Ahvaz, Iran.
Abstract:   (1368 Views)
Aim and Background: RNA isolation is difficult in plant that contain large amounts of polysaccharides and polyphenol compounds that bind and co-precipitate with RNA. Therefore, in this study eleven different methods were evaluated to extract RNA from leaves of apple rich in phenolic compounds. Material and methods:The quantity and quality of extracted RNA were determined by 1.2% nano-drop spectrophotometric at absorbance of 230, 260 and 280nm, agarose gel electrophoresis and cDNA synthesis and RT-PCR analysis by specific primer for apple Actin were performed . Results:The results indicated that methods of Djami-Tchatchou et al., (2012) and Rajakani et al.,(2013) were the most suitable methods for RNA extraction from apple leaves. The yield of total RNA extracted by these methods varies from 753.6 to 800.2 and 362.1 to 452.6 ng/µl in apple leaves , respectively. Distinct bands of 28S and 18S ribosomal RNA were observed in electrophoresis Discuss: In two protocols, ratios of A260/A280 and A260/A230 ranged from 1.99 to 2.03 and from 2.07 to 2.17, respectively This is indicative of a high RNA purity and the absence of contamination with polyphenolic, protein and polysaccharide compounds Conclusion: The high quality for RT-PCR reaction by two methods showed the extracted RNA can be used with assurance in downstream molecular techniques.
Keywords: Priming, Salinity Stress, Germination, Cucumber
Full-Text [PDF 370 kb]   (1152 Downloads)    
Type of Study: Research Article | Subject: Genetics
Received: 2017/10/24 | Accepted: 2017/10/24 | Published: 2017/10/24
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Montazeri S, Sadat S. Comparative study of different protocols for isolation of RNA from apple leaves (Malus domestica). NCMBJ. 2017; 7 (27) :55-62
URL: http://ncmbjpiau.ir/article-1-1001-en.html

Volume 7, Issue 27 (9-2017) Back to browse issues page
مجله تازه های بیوتکنولوژی سلولی - مولکولی New Cellular and Molecular Biotechnology Journal
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