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Simple and Efficient Protocol for RNA and DNA extraction from Rice (Oryza sativa L.) for Downstream Applications

Author Affiliations

  • 1Centre for Advanced Studies in Plant Biotechnology and Genetic Engineering, Department of Biosciences, Saurashtra University, Rajkot, Gujarat,

Int. Res. J. Biological Sci., Volume 4, Issue (2), Pages 62-67, February,10 (2015)

Abstract

Plant DNA and RNA extraction is difficult due to the presence of metabolites that interfere with isolation procedures and downstream applications. In the present study DNA and RNA extraction protocol standardized from rice (Oryza sativa L.). Isolation of DNA using Triton-X-100 based extraction method with PVPP treatment efficiently removes metabolites and yield with high quality DNA from rice coleoptiles.RNA isolation methods based on TRIzol was suitable for rice leaf whereas in seed, TrisHCl (pH 9.0) with mercaptoethanol protocol gave best results. The isolated DNA and RNA proved to be suitable for PCR and RT-PCR amplification, respectively. The technique is reproducible and can be applied for PCR based markers identification and gene expression studies.

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