Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.18/3775
Título: Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
Autor: Churro, Catarina
Valério, Elisabete
Pereira, Paulo
Vasconcelos, Vitor
Palavras-chave: Cyanobacteria
Glutaraldehyde
Formaldehyde
Lugol’s iodine
Real-time qPCR
Planktothrix
Água e Solo
Data: 2015
Editora: Asociación Ibérica de Limnología
Citação: Limnetica. 2015;34(1):173-86
Resumo: The study and monitoring of cyanobacterial blooms often involves the use of preserved samples to avoid cellular degradation. However, preserved samples may not be suitable for molecular biology studies because preservation methods can interfere with DNA quality/quantity. Real-time quantitative PCR analysis (qPCR) has been widely applied in molecular analysis and is considered a promising method for monitoring purposes. This study intended to evaluate the applicability of the real-time qPCR technique in samples that were subjected to different methods of preservation: (1) 15% Lugol’s iodine solution (2) 4% formaldehyde and (3) 25% glutaraldehyde. The ability to amplify and quantify DNA extracted from Planktothrix agardhii was assessed using the rpoC1 gene as the target fragment in both raw water samples and in vitro cultures. No reliable DNA amplification was obtained from glutaraldehyde-preserved samples. Successful amplification was obtained from Lugol’s and formaldehyde-preserved samples. In this case, however, the quantification that was achieved by real-time PCR cannot be used to infer cell numbers, because the Ct values that were obtained from the Lugol’s and formaldehydepreserved samples were significantly higher than the Ct values that were obtained from the unpreserved samples. Therefore real-time PCR can be used for the detection and identification of cyanobacteria in preserved samples but no reliable cell quantification can be performed using this method.
Descrição: Publicaciones del III Congreso Ibérico de Cianotoxinas (Blanes, 2013)
Peer review: yes
URI: http://hdl.handle.net/10400.18/3775
ISSN: 0213-8409
Versão do Editor: http://www.limnetica.com/Limnetica/Limne34/L34a173_qPCR_preserved_cyanobacteria_samples.pdf
Aparece nas colecções:DSA - Artigos em revistas internacionais

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