Genes that Contribute to the Survival and Virulence of Dickeya Dadantii

Genes that Contribute to the Survival and Virulence of Dickeya Dadantii

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RNA integrity is critical for successful RNA quantitation and the level of integrity required for quantitation has not been determined for bacterial RNA. Three RNA isolation methods were evaluated for their ability to produce high-quality RNA from D. dadantii. A hot SDS-hot phenol RNA method gave the highest RNA quality and required only two DNase treatments to remove contaminating DNA. The assessment of RNA integrity was critical for obtaining meaningful gene expression data. RIN values below 7.0 resulted in high variation and loss of statistical significance when gene expression was analyzed by real-time qRT-PCR. This work provides guidelines for RNA isolation and quality assessment that will be valuable for gene expression studies in a wide range of bacteria.Contaminating DNA remained a problem with the hot SDS-Trizol method; RNA samples required repeated, rigorous DNase ... The assessment of RNA integrity using the Agilent 2100 BioAnalyzer was critical for obtaining meaningful geneanbsp;...


Title:Genes that Contribute to the Survival and Virulence of Dickeya Dadantii
Author: Courtney Elaine Jahn
Publisher:ProQuest - 2008
ISBN-13:

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