New design strategy for development of specific primer sets for PCR-based detection of Chlorophyceae and Bacillariophyceae in environmental samples.
Studying aquatic microalgae is essential for monitoring biodiversity and water quality. We designed new sets of 18S rRNA PCR primers for Chlorophyceae and Bacillariophyceae by using the ARB software and implementing a virtual PCR program. The results of specificity analysis showed that most of the targeted algal families were identified and nontargeted organisms, such as fungi or ciliates, were excluded. These newly developed PCR primer sets were also able to amplify microalgal rRNA genes from environmental samples with accurate specificity. These tools could be of great interest for studying freshwater microalgal ecology and for developing bioindicators of the health status of aquatic environments.
Moro
Claire Valiente
CV
UMR 6023 Laboratoire Microorganismes: Génome et Environnement (CNRS-Université Blaise Pascal, Clermont-Ferrand II), 63177 Aubière, France. valienteclaire@yahoo.fr
Crouzet
Olivier
O
Rasconi
Séréna
S
Thouvenot
Antoine
A
Coffe
Gérard
G
Batisson
Isabelle
I
Bohatier
Jacques
J
D000460:Chlorophyta / Q000235:genetics / Q000302:isolation & purification*; D019295:Computational Biology; D003198:Computer Simulation; D017931:DNA Primers / Q000235:genetics*; D048028:DNA, Algal / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D017377:Diatoms / Q000235:genetics / Q000302:isolation & purification*; D004783:Environmental Microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D048030:RNA, Algal / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D017931:DNA Primers; D048028:DNA, Algal; D004275:DNA, Ribosomal; D048030:RNA, Algal; D012337:RNA, Ribosomal, 18S