Extraction of High Quality of RNA and Construction of a Suppression Subtractive Hybridization (SSH) Library from Chestnut Rose
(Rosa roxburghii Tratt) |
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Authors: | Qiang Xu Xiaopeng Wen Nengguo Tao Zhiyong Hu Hailin Yue Xiuxin Deng |
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Institution: | (1) National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, 430070, Wuhan, P.R. China;(2) Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou Univeristy, Guiyang, P.R. China |
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Abstract: | Chestnut rose (Rosa roxburghii Tratt) is a rare fruit crop of promising economical importance in fruit and ornamental exploitation in China. Isolation of
high quality RNA from chestnut rose is difficult due to its high levels of polyphenols, polysaccharides and other compounds,
but a modified CTAB extraction procedure without phenol gave satisfactory results. High concentrations of PVP (2%, w/v), CTAB
(2%, w/v) and β-mercaptoethanol (4%, v/v) were used in the extraction buffer to improve RNA quality. The average yield was about 200 μg RNA g−1 fresh leaves. The isolated RNA was of sufficient quality for construction of suppression subtraction hybridization (SSH)
library, which allowed the isolation of several pathogen-induced defense genes.
Qiang Xu and Xiaopeng Wen - Contribute to this work equally
Revisions requested 3 November 2005; Revisions received 18 January 2006 |
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Keywords: | chestnut rose RNA isolation Rosa roxburghii Tratt suppression subtractive hybridization library |
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