Publication
Applicability of the real-time PCR assay in the ampliļ¬cation of cyanobacterial DNA from preserved samples
| dc.contributor.author | Churro, Catarina | |
| dc.contributor.author | ValƩrio, Elisabete | |
| dc.contributor.author | Pereira, Paulo | |
| dc.contributor.author | Vasconcelos, Vitor | |
| dc.date.accessioned | 2016-04-06T11:43:19Z | |
| dc.date.available | 2016-04-06T11:43:19Z | |
| dc.date.issued | 2015 | |
| dc.description | Publicaciones del III Congreso IbƩrico de Cianotoxinas (Blanes, 2013) | pt_PT |
| dc.description.abstract | 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% Lugolās 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 ampliļ¬cation was obtained from glutaraldehyde-preserved samples. Successful ampliļ¬cation was obtained from Lugolās and formaldehyde-preserved samples. In this case, however, the quantiļ¬cation that was achieved by real-time PCR cannot be used to infer cell numbers, because the Ct values that were obtained from the Lugolās and formaldehydepreserved samples were signiļ¬cantly higher than the Ct values that were obtained from the unpreserved samples. Therefore real-time PCR can be used for the detection and identiļ¬cation of cyanobacteria in preserved samples but no reliable cell quantiļ¬cation can be performed using this method. | pt_PT |
| dc.description.sponsorship | This research was supported by the FundaĆ§Ć£o para CiĆŖncia e Tecnologia, Portugal (FCT) through the project PPCDT/AMB/67075/2006. The authors also acknowledge the PhD research grant SFRH/BD65706/2009 to C. Churro and the Post-Doc research grant SFRH/BPD/75922/2011 to E. ValĆ©rio. | pt_PT |
| dc.identifier.citation | Limnetica. 2015;34(1):173-86 | pt_PT |
| dc.identifier.issn | 0213-8409 | |
| dc.identifier.uri | http://hdl.handle.net/10400.18/3775 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.publisher | AsociaciĆ³n IbĆ©rica de LimnologĆa | pt_PT |
| dc.relation | The application of quantitative real-time PCR to studies of the abundance and toxicity of cyanobacteria in Portuguese potable and recreational water supplies | |
| dc.relation.publisherversion | http://www.limnetica.com/Limnetica/Limne34/L34a173_qPCR_preserved_cyanobacteria_samples.pdf | pt_PT |
| dc.subject | Cyanobacteria | pt_PT |
| dc.subject | Glutaraldehyde | pt_PT |
| dc.subject | Formaldehyde | pt_PT |
| dc.subject | Lugolās iodine | pt_PT |
| dc.subject | Real-time qPCR | pt_PT |
| dc.subject | Planktothrix | pt_PT |
| dc.subject | Ćgua e Solo | pt_PT |
| dc.title | Applicability of the real-time PCR assay in the ampliļ¬cation of cyanobacterial DNA from preserved samples | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.awardTitle | The application of quantitative real-time PCR to studies of the abundance and toxicity of cyanobacteria in Portuguese potable and recreational water supplies | |
| oaire.awardURI | info:eu-repo/grantAgreement/FCT/5876-PPCDTI/PTDC%2FAMB%2F67075%2F2006/PT | |
| oaire.citation.conferencePlace | Madrid, Espanha | pt_PT |
| oaire.citation.endPage | 186 | pt_PT |
| oaire.citation.startPage | 173 | pt_PT |
| oaire.citation.title | Limnetica | pt_PT |
| oaire.citation.volume | 34(1) | pt_PT |
| oaire.fundingStream | 5876-PPCDTI | |
| project.funder.identifier | http://doi.org/10.13039/501100001871 | |
| project.funder.name | FundaĆ§Ć£o para a CiĆŖncia e a Tecnologia | |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | article | pt_PT |
| relation.isProjectOfPublication | 08dd2d8c-1680-4e89-9750-7c8987aa1a2e | |
| relation.isProjectOfPublication.latestForDiscovery | 08dd2d8c-1680-4e89-9750-7c8987aa1a2e |