NDLI: Evolution of the MIDTAL microarray: the adaption and testing of oligonucleotide 18S and 28S rDNA probes and evaluation of subsequent microarray generations with Prymnesium spp. cultures and field samples.

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Evolution of the MIDTAL microarray: the adaption and testing of oligonucleotide 18S and 28S rDNA probes and evaluation of subsequent microarray generations with Prymnesium spp. cultures and field samples.

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Evolution of the MIDTAL microarray: the adaption and testing of oligonucleotide 18S and 28S rDNA probes and evaluation of subsequent microarray generations with Prymnesium spp. cultures and field samples.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Touzet, Nicolas Raine, Robin Fleming, Gerard T. A. McCoy, Gary R.
Spatial Coverage	Ireland France
Description	Author Affiliation: McCoy GR ( The Ryan Institute, National University of Ireland, Galway, Martin Ryan Building, Galway, Ireland, gary.mccoy8@gmail.com.)
Abstract	The toxic microalgal species Prymnesium parvum and Prymnesium polylepis are responsible for numerous fish kills causing economic stress on the aquaculture industry and, through the consumption of contaminated shellfish, can potentially impact on human health. Monitoring of toxic phytoplankton is traditionally carried out by light microscopy. However, molecular methods of identification and quantification are becoming more common place. This study documents the optimisation of the novel Microarrays for the Detection of Toxic Algae (MIDTAL) microarray from its initial stages to the final commercial version now available from Microbia Environnement (France). Existing oligonucleotide probes used in whole-cell fluorescent in situ hybridisation (FISH) for Prymnesium species from higher group probes to species-level probes were adapted and tested on the first-generation microarray. The combination and interaction of numerous other probes specific for a whole range of phytoplankton taxa also spotted on the chip surface caused high cross reactivity, resulting in false-positive results on the microarray. The probe sequences were extended for the subsequent second-generation microarray, and further adaptations of the hybridisation protocol and incubation temperatures significantly reduced false-positive readings from the first to the second-generation chip, thereby increasing the specificity of the MIDTAL microarray. Additional refinement of the subsequent third-generation microarray protocols with the addition of a poly-T amino linker to the 5' end of each probe further enhanced the microarray performance but also highlighted the importance of optimising RNA labelling efficiency when testing with natural seawater samples from Killary Harbour, Ireland.
ISSN	09441344
Issue Number	13
Volume Number	22
e-ISSN	16147499
Journal	Environmental Science and Pollution Research
Language	English
Publisher	Springer
Publisher Date	2015-07-01
Publisher Place	Germany
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Environmental Monitoring Methods Haptophyta Genetics Oligonucleotide Array Sequence Analysis Oligonucleotide Probes Acclimatization Dna, Ribosomal France Classification Humans In Situ Hybridization, Fluorescence Ireland Microalgae Nucleic Acid Hybridization Oligonucleotides Phytoplankton Rna Seawater Journal Article Research Support, Non-u.s. Gov't Discipline Environmental Science Discipline Environmental Chemistry
Content Type	Text
Resource Type	Article
Subject	Environmental Chemistry Health, Toxicology and Mutagenesis Pollution Medicine

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