Isolation of functional total RNA from Tilia cordata leaves and pollen

Ognjenovic, Jana and Tantoush, Ziyad Omar and Jankov, Ratko M and Ćirković-Veličković, Tanja and Vukmirica, Jelena (2012) Isolation of functional total RNA from Tilia cordata leaves and pollen. Journal of the Serbian Chemical Society, 77 (8). pp. 1003-1012. ISSN 0352-5139 (FP7- 256716)

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Abstract

The conditions required for the isolation of high quality total RNA from European linden (Tilia cordata) leaves and pollen were determined. Pure total RNA was isolated from linden leaves utilizing a Qiagen plant mini kit, while the total RNA isolated from linden pollen using this method was degraded. Successful isolation of total RNA from both linden pollen and leaves, however, was achieved following TRIzol™ preparation of the total RNA. The total RNA isolated using TRIzol™ was contaminated with genomic DNA but treatment with the enzyme DNase, in solution or on-column, efficiently removed the genomic DNA. Furthermore, the conditions for the elimination of genomic DNA contamination on-column and isolation of pure total RNA from leaves were optimized. The isolated total RNA from both leaves and pollen was used successfully in first- and second-strand cDNA synthesis reactions and in a reverse transcription polymerase chain reaction (RT-PCR), demonstrating that the total RNA isolated using this method was functional. In conclusion, pure and functional total RNA from T. cordata leaves and pollen (27.8±7.9 μg g-1 leaves; 25.7±1.1 μg g-1 pollen) could be obtained and was suitable for application in further molecular biology studies.

Keywords

Tilia cordata; leaves; pollen; total RNA; RT-PCR; cDNA

Item Type:	Article
FP7 Grant Agreement Number:	256716
FrameWork Programmes:	SP4-Capacities
Scientific Areas:	Research Potential
Contact Email Address:	jognjenovic@chem.bg.ac.rs
Last Modified:	26 Sep 2012 10:06
Access rights:	Open access
Output type:	Article
URI:	http://eprints.kobson.nb.rs/id/eprint/38

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