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| Reference : Novel FISH and quantitative PCR protocols to monitor artificial consortia composed of di... |
| Scientific journals : Article | |||
| Life sciences : Microbiology | |||
| http://hdl.handle.net/2268/91934 | |||
| Novel FISH and quantitative PCR protocols to monitor artificial consortia composed of different hydrogen-producing Clostridium spp. | |
| English | |
| [fr] Protocoles nouveaux de FISH et PCR quantitative pour suivre un consortium artificiel de clostridies productrices d'hydrogène | |
Savichtcheva, Olga  [ > > ] | |
| Joris, Bernard [Université de Liège - ULg > Département des sciences de la vie > Physiologie et génétique bactériennes >] | |
| Wilmotte, Annick [Université de Liège - ULg > Département des sciences de la vie > Physiologie et génétique bactériennes >] | |
| Calusinska, Magdalena [Université de Liège - ULg > > Centre d'ingénierie des protéines >] | |
| Apr-2011 | |
| International Journal of Hydrogen Energy | |
| Pergamon Press - An Imprint of Elsevier Science | |
| 36 | |
| Hysydays | |
| 7530-7542 | |
| International | |
| 0360-3199 | |
| [en] Fluorescent in situ hybridisation ; Artificial consortium ; Quantitative PCR | |
| [en] The use of an artificial consortium composed of selected hydrogen-producing species,
instead of a natural anaerobic sludge, has been proposed for biohydrogen production. In order to monitor such a consortium composed of different Clostridium spp., new protocols were tested for two different assays, FISH and qPCR. New species-specific FISH probes and qPCR primer sets were developed and optimised for three strains: Clostridium butyricum, Clostridium felsineum and Clostridium pasteurianum, that were used in a consortium. Application of a fast two-step FISH protocol, with pre-treatment step at 90 C for 5 min and a subsequent hybridisation step at higher temperature (55 C) for 20 min resulted in a much shorter analytical time compared to the standard FISH procedure (46 C for 2e3 h) and gave a high hybridisation performance. Moreover, to accurately quantify each microorganism by qPCR assay, two innovations were tested: the direct use of cell lysates (omitting the DNA extraction step) and the use of two alternative molecular markers, recA and gyrA. These markers are present in single copies in the genome, whereas there are multiple copies of the ribosomal operons. This resulted in the development of accurate, reliable and fast FISH and qPCR assays for routine monitoring of the dynamics of artificial hydrogen-producing microbial consortia. Moreover, both techniques can be easily adapted to new Clostridium strains. | |
| Centre d'Ingénierie des Protéines - CIP | |
| Communauté française de Belgique - CfB | |
| Microbiological production of hydrogen: study of microalgal and bacterial processes | |
| Researchers | |
| http://hdl.handle.net/2268/91934 | |
| 10.1016/j.ijhydene.2011.03.097 | |
| http://www.sciencedirect.com/science/article/pii/S0360319911006872 | |
| This article appeared in a journal published by Elsevier. The attached
copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues. Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. In most cases authors are permitted to post their version of the article (e.g. in Word or Tex form) to their personal website or institutional repository. |
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