A method for extracting a high-quality RNA from Symbiodinium sp.

Rosic, N. N. and Hoegh-Guldberg, O (2009) A method for extracting a high-quality RNA from Symbiodinium sp.. Journal of Applied Phycology, 22 2: 139-146.


Author Rosic, N. N.
Hoegh-Guldberg, O
Title A method for extracting a high-quality RNA from Symbiodinium sp.
Formatted title A method for extracting a high-quality RNA from Symbiodinium sp.
Journal name Journal of Applied Phycology   Check publisher's open access policy
ISSN 0921-8971
Publication date 2009-04
Year available 2009
Sub-type Article (original research)
DOI 10.1007/s10811-009-9433-x
Volume 22
Issue 2
Start page 139
End page 146
Total pages 8
Editor Michael A Borowitzka
Place of publication Netherlands
Publisher Springer - Netherlands
Collection year 2010
Language eng
Subject 060405 Gene Expression (incl. Microarray and other genome-wide approaches)
8298 Environmentally Sustainable Plant Production
C1
Abstract Good quality RNA is essential for a range of analyses including microarray and gene expression studies. A number of methods for RNA extraction from symbiotic dinoflagellates were assessed for their ability to recover a high-quality RNA applicable for evaluation of gene expression profiles. The recovery and quality of the obtained RNA were evaluated with respect to UV light absorbance profiles and automated microcapillary electrophoretic RNA separation. A modified RNA extraction procedure that combines two existing commercial kits, Trizol and Qiagen RNeasy kits, was efficiently employed for the recovery of a high-quality RNA under specific homogenization conditions. Cell homogenization using glass beads at the speed of 4,500 rpm for up to 6 min resulted in a good RNA recovery and preserved RNA integrity. A high-quality RNA obtained following the described procedure was successfully applied in reverse transcriptase-polymerase chain reaction (PCR) and in quantitative PCR studies. Gene expression profiles were changed with RNA extraction procedure, with the highest transcript numbers at precise conditions of cell homogenization. RNA samples with RNA integrity number values from 6 and above were recommended for downstream applications. This sequence of RNA isolation and RNA evaluation represents a methodological improvement required for functional genomic studies in dinoflagellates.
Keyword Dinoflagellate
Symbiodinium
RNA
Real-time PCR
Gene expression
Bioanalyzer
MICROCAPILLARY ELECTROPHORESIS TRACES
REEF-BUILDING CORALS
GENE-EXPRESSION
STRESS CAUSES
DINOFLAGELLATE
MICROADRIATICUM
INTEGRITY
IMPACT
Q-Index Code C1
Q-Index Status Confirmed Code

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
Centre for Marine Studies Publications
 
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Created: Sun, 28 Mar 2010, 00:04:17 EST