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Polyethylene glycol and polyvinylpirrolidone effect on bacterial rRNA extraction and hybridization from cells exposed to tannins

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dc.creator Arcuri Pedro Braga
dc.creator Thonney Michael Larry
dc.creator Schofield Peter
dc.creator Pell Alice Nelson
dc.date 2003
dc.date.accessioned 2013-06-01T09:35:07Z
dc.date.available 2013-06-01T09:35:07Z
dc.date.issued 2013-06-01
dc.identifier http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2003000900008
dc.identifier http://www.doaj.org/doaj?func=openurl&genre=article&issn=0100204X&date=2003&volume=38&issue=9&spage=1073
dc.identifier.uri http://koha.mediu.edu.my:8181/jspui/handle/123456789/7869
dc.description In order to detect fluctuations in ruminal microbial populations due to forage tannins using 16S ribosomal RNA (rRNA) probes, recovery of intact rRNA is required. The objective of this work was to evaluate the effect of polyethylene glycol (PEG) and polyvinylpirrolidone (PVP) on extraction of bacterial rRNA, in the presence of tannins from tropical legume forages and other sources, that hybridize with oligonucleotide probes. Ruminococcus albus 8 cells were exposed to 8 g/L tannic acid or 1 g/L condensed tannins extracted from Acacia angustissima, banana (Musa sp.) skin, Desmodium ovalifolium, red grape (Vitis vinifera) skin and Inga edulis, or no tannins. Cells were rinsed with Tris buffer pH 7 containing either 8% PEG or 6% PVP prior to cell lysis. Total RNA samples rinsed with either PEG or PVP migrated through denaturing agarose gels. The 16S rRNA bands successfully hybridized with a R. albus species-specific oligonucleotide probe, regardless of tannin source. The effect of rinsing buffers on the density of 16S rRNA bands, as well as on the hybridization signals was compared. There were significant effects (P<0.01) when the controls were compared to either buffer treatments due to tannin type, buffer used and the interaction of tannin type and buffer. The significant interaction indicates the influence of tannin type on the parameters evaluated.
dc.publisher Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
dc.source Pesquisa Agropecuária Brasileira
dc.subject Ruminococcus albus
dc.subject microbial ecology
dc.subject animal nutrition
dc.title Polyethylene glycol and polyvinylpirrolidone effect on bacterial rRNA extraction and hybridization from cells exposed to tannins


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