A micromethod for high throughput RNA extraction in forest trees
| Autor: | Raúl Herrera, Franck Salin, Jorge A. P. Paiva, Christophe Plomion, Philippe Chaumeil, Grégoire Le Provost |
|---|---|
| Přispěvatelé: | Biodiversité, Gènes & Communautés (BioGeCo), Institut National de la Recherche Agronomique (INRA)-Université de Bordeaux (UB), Universidad de Talca |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
0106 biological sciences
arbre forestier DNA Complementary pinus pin [SDV.BC]Life Sciences [q-bio]/Cellular Biology Biology Bioinformatics 01 natural sciences General Biochemistry Genetics and Molecular Biology Trees 03 medical and health sciences chemistry.chemical_compound Gene Expression Regulation Plant Gene expression microméthode lcsh:QH301-705.5 ComputingMilieux_MISCELLANEOUS 030304 developmental biology Regulation of gene expression 0303 health sciences Chromatography Reverse Transcriptase Polymerase Chain Reaction Extraction (chemistry) Reproducibility of Results RNA CDNA Library Construction General Medicine 15. Life on land Microarray Analysis RNA extraction Reverse transcription polymerase chain reaction micromethod arn t Genetic Techniques chemistry lcsh:Biology (General) RNA Plant Cetrimonium Compounds extraction General Agricultural and Biological Sciences DNA forest trees 010606 plant biology & botany |
| Zdroj: | Biological Research, Vol 40, Iss 3, Pp 291-297 (2007) Biological Research, Volume: 40, Issue: 3, Pages: 291-297, Published: 2007 Biological Research Biological Research, Sociedad de Biología de Chile, 2007, 40 (3), pp.291-297 Biological Research 3 (40), 291-297. (2007) CIÊNCIAVITAE Scopus-Elsevier Biological Research v.40 n.3 2007 SciELO Chile CONICYT Chile instacron:CONICYT HAL |
| ISSN: | 0717-6287 0716-9760 |
| Popis: | A large quantity of high quality RNA is often required in the analysis of gene expression. However, RNA extraction from samples taken from woody plants is generally complex, and represents the main limitation to study gene expression, particularly in refractory species like conifers. Standard RNA extraction protocols are available but they are highly time consuming, and not adapted to large scale extraction. Here we present a high-throughput RNA extraction protocol. This protocol was adapted to a micro-scale by modifying the classical cetyltrimethylammonium (CTAB) protocol developed for pine: (i) quantity of material used (100-200 mg of sample), (ii) disruption of samples in microtube using a mechanical tissue disrupter, and (iii) the use of SSTE buffer. One hundred samples of woody plant tissues/organs can be easily treated in two working days. An average of 15 \ig of high quality RNA per sample was obtained. The RNA extracted is suitable for applications such as real time reverse transcription polymerase chain reaction, cDNA library construction or synthesis of complex targets for microarray analysis. |
| Databáze: | OpenAIRE |
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