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An RNA isolation system for plant tissues rich in secondary metabolites

IR@CSIR-IHBT

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Title An RNA isolation system for plant tissues rich in
secondary metabolites
 
Creator Ghawana, S
Paul, A
Kumar, Hitesh
Kumar, A
Singh, H
Bhardwaj , P K
Rani, A
Singh, R S
Raizada, J
Singh, K
Kumar , Sanjay
 
Subject Plant Biotechnology
 
Description Background: Secondary metabolites are reported to interfere with the isolation of RNA particularly with the
recipes that use guanidinium-based salt. Such interference was observed in isolation of RNA with medicinal plants
rheum (Rheum australe) and arnebia (Arnebia euchroma). A rapid and less cumbersome system for isolation of RNA
was essential to facilitate any study related to gene expression.
Findings: An RNA isolation system free of guanidinium salt was developed that successfully isolated RNA from
rheum and arnebia. The method took about 45 min and was successfully evaluated on twenty one tissues with
varied secondary metabolites. The A260/280 ratio ranged between 1.8 - 2.0 with distinct 28 S and 18 S rRNA bands
visible on a formaldehyde-agarose gel.
Conclusions: The present manuscript describes a rapid protocol for isolation of RNA, which works well with all the
tissues examined so far. The remarkable feature was the success in isolation of RNA with those tissues, wherein the
most commonly used methods failed. Isolated RNA was amenable to downstream applications such as reverse
transcription-polymerase chain reaction (RT-PCR), differential display (DD), suppression subtractive hybridization
(SSH) library construction, and northern hybridization.
 
Date 2011
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/613/1/506_2011_9-12-2011.pdf
Ghawana, S and Paul, A and Kumar, Hitesh and Kumar, A and Singh, H and Bhardwaj , P K and Rani, A and Singh, R S and Raizada, J and Singh, K and Kumar , Sanjay (2011) An RNA isolation system for plant tissues rich in secondary metabolites. BMC Research Notes, 4. p. 85.
 
Relation http://ihbt.csircentral.net/613/