An RNA isolation system for plant tissues rich in secondary metabolites

被引：211

作者：

Ghawana S. ^{[1
,2
]}

Paul A. ^{[1
]}

Kumar H. ^{[1
,3
]}

Kumar A. ^{[1
]}

Singh H. ^{[1
,4
]}

Bhardwaj P.K. ^{[1
,5
]}

Rani A. ^{[1
,6
]}

Singh R.S. ^{[1
]}

Raizada J. ^{[1
]}

Singh K. ^{[1
,7
]}

Kumar S. ^{[1
]}

机构：

[1] Biotechnology Division, Institute of Himalayan Bioresource Technology (CSIR), Palampur-176 061, Himachal Pradesh

[2] National Institute of Plant Genome Research, New Delhi- 110 067, Aruna Asaf Ali Marg

[3] Department of Botany, SCVB Government College, Palampur-176 061, Himachal Pradesh

[4] Biotechnology Division, Lyallpur Khalsa College, Jalandhar-144 001, Punjab

[5] Regional Centre of Institute of Bioresources and Sustainable Development (DBT), Tadong-737 102, Sikkim

[6] Vittal Mallya Scientific Research Foundation, Bangalore-560 076, Karnataka

[7] Department of Biotechnology, Panjab University

来源：

BMC Research Notes | / 4卷 / 1期

关键词：

Secondary Metabolite; Suppression Subtractive Hybridization; Differential Display; NaOAc; Downstream Application;

D O I：

10.1186/1756-0500-4-85

中图分类号：

学科分类号：

摘要：

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 A260/280ratio 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. © 2011 Kumar et al; licensee BioMed Central Ltd.

引用

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