A Protocol to Remove Colored Metabolites and Other Inhibitors from Plant Tissues to Facilitate RNA Isolation Suitable for Downstream Applications
IR@IHBT: CSIR-Institute of Himalayan Bioresource Technology, Palampur
View Archive Info| Field | Value | |
| Title |
A Protocol to Remove Colored Metabolites and Other Inhibitors from Plant
Tissues to Facilitate RNA Isolation Suitable for Downstream Applications
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| Creator |
Singh, Ravi S
Kumar , Sanjay |
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| Subject |
Plant sciences
Plant Biotechnology |
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| Description |
This work developed a protocol to remove colored metabolites and other interfering substances
to facilitate RNA isolation. These metabolites otherwise hinder RNA isolation and
downstream applications. The developed protocol used sodium dodecyl sulphate, ethylenediamine
tetra-acetic acid, and ethanol in a definite ratio that removed the said metabolites
from the tissue and aided isolation of RNA using the existing methods. The protocol was
developed for red colored roots of Arnebia euchroma and was extended to other colored tissues
[rhizome of Rheum australe (Himalayan Rhubarb) and taproot of Daucus carota (purple
carrot)] with success. Without inclusion of our protocol, the existing methods could not
isolate good quality RNA from these tissues. RNA isolated by the developed protocol had
A260/280 ratio of 1.88–1.93, A260/230 ratio of 1.94–2.0, and RNA integrity number of 6.3–8.0.
RNA was amenable to downstream applications such as reverse-transcription polymerase
chain reaction and primer extension assay.
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| Date |
2012
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://ihbt.csircentral.net/1339/1/8_A_Protocol.pdf
Singh, Ravi S and Kumar , Sanjay (2012) A Protocol to Remove Colored Metabolites and Other Inhibitors from Plant Tissues to Facilitate RNA Isolation Suitable for Downstream Applications. Journal of Biotechnology Progress, 28. pp. 1303-1307. |
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| Relation |
http://ihbt.csircentral.net/1339/
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