Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria.
Cyanobacteria are a group of photosynthetic prokaryotes that have a diverse morphology, minimal nutritional requirements and metabolic plasticity that has made them attractive organisms to use in biotechnological applications. The use of these organisms as cell factories requires the knowledge of th...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3319621?pdf=render |
| _version_ | 1819041077762457600 |
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| author | Filipe Pinto Catarina C Pacheco Daniela Ferreira Pedro Moradas-Ferreira Paula Tamagnini |
| author_facet | Filipe Pinto Catarina C Pacheco Daniela Ferreira Pedro Moradas-Ferreira Paula Tamagnini |
| author_sort | Filipe Pinto |
| collection | DOAJ |
| description | Cyanobacteria are a group of photosynthetic prokaryotes that have a diverse morphology, minimal nutritional requirements and metabolic plasticity that has made them attractive organisms to use in biotechnological applications. The use of these organisms as cell factories requires the knowledge of their physiology and metabolism at a systems level. For the quantification of gene transcripts real-time quantitative polymerase chain reaction (RT-qPCR) is the standard technique. However, to obtain reliable RT-qPCR results the use and validation of reference genes is mandatory. Towards this goal we have selected and analyzed twelve candidate reference genes from three morphologically distinct cyanobacteria grown under routinely used laboratory conditions. The six genes exhibiting less variation in each organism were evaluated in terms of their expression stability using geNorm, NormFinder and BestKeeper. In addition, the minimum number of reference genes required for normalization was determined. Based on the three algorithms, we provide a list of genes for cyanobacterial RT-qPCR data normalization. To our knowledge, this is the first work on the validation of reference genes for cyanobacteria constituting a valuable starting point for future works. |
| first_indexed | 2024-12-21T09:19:16Z |
| format | Article |
| id | doaj.art-03299acd91f644cdba750214814d588a |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T09:19:16Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-03299acd91f644cdba750214814d588a2022-12-21T19:09:04ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0174e3498310.1371/journal.pone.0034983Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria.Filipe PintoCatarina C PachecoDaniela FerreiraPedro Moradas-FerreiraPaula TamagniniCyanobacteria are a group of photosynthetic prokaryotes that have a diverse morphology, minimal nutritional requirements and metabolic plasticity that has made them attractive organisms to use in biotechnological applications. The use of these organisms as cell factories requires the knowledge of their physiology and metabolism at a systems level. For the quantification of gene transcripts real-time quantitative polymerase chain reaction (RT-qPCR) is the standard technique. However, to obtain reliable RT-qPCR results the use and validation of reference genes is mandatory. Towards this goal we have selected and analyzed twelve candidate reference genes from three morphologically distinct cyanobacteria grown under routinely used laboratory conditions. The six genes exhibiting less variation in each organism were evaluated in terms of their expression stability using geNorm, NormFinder and BestKeeper. In addition, the minimum number of reference genes required for normalization was determined. Based on the three algorithms, we provide a list of genes for cyanobacterial RT-qPCR data normalization. To our knowledge, this is the first work on the validation of reference genes for cyanobacteria constituting a valuable starting point for future works.http://europepmc.org/articles/PMC3319621?pdf=render |
| spellingShingle | Filipe Pinto Catarina C Pacheco Daniela Ferreira Pedro Moradas-Ferreira Paula Tamagnini Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria. PLoS ONE |
| title | Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria. |
| title_full | Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria. |
| title_fullStr | Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria. |
| title_full_unstemmed | Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria. |
| title_short | Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria. |
| title_sort | selection of suitable reference genes for rt qpcr analyses in cyanobacteria |
| url | http://europepmc.org/articles/PMC3319621?pdf=render |
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