Development of cPCR technique for detection and enumeration of Prevotella bryantii
Competitive PCR (cPCR) system for the detection and enumeration of Prevotella bryantii cells in rumen samples was developed. PBB14 primer, specific for P. bryantii was used as the reverse PCR primer and EUB 338 bacterial primer as the forward primer. An internal DNA control, containing primer specif...
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| Format: | Article |
| Language: | English |
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University of Ljubljana Press (Založba Univerze v Ljubljani)
1999-12-01
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| Series: | Acta Agriculturae Slovenica |
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| Online Access: | https://journals.uni-lj.si/aas/article/view/15907 |
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| _version_ | 1849772101330796544 |
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| author | Katarina TEPŠIČ Gorazd AVGUŠTIN |
| author_facet | Katarina TEPŠIČ Gorazd AVGUŠTIN |
| author_sort | Katarina TEPŠIČ |
| collection | DOAJ |
| description | Competitive PCR (cPCR) system for the detection and enumeration of Prevotella bryantii cells in rumen samples was developed. PBB14 primer, specific for P. bryantii was used as the reverse PCR primer and EUB 338 bacterial primer as the forward primer. An internal DNA control, containing primer specific sequences at 5’ and 3’ ends, but lacking 41 bp at the middle of the sequence, was constructed. C-PCR products were quantified by capillary electrophoresis and the results were calculated with regression equation (Reilly and Attwood, 1998). By coamplification of P.bryantii B14 genomic DNA extracted from known numbers of cells and internal control, standard curve was constructed which enables quantification of P.bryantii cells in the range of 2.15 x 103 to 4.3 x 104 cells. |
| format | Article |
| id | doaj-art-68c8b73e40814e4bb4eb211be8547f74 |
| institution | DOAJ |
| issn | 1854-1941 |
| language | English |
| publishDate | 1999-12-01 |
| publisher | University of Ljubljana Press (Založba Univerze v Ljubljani) |
| record_format | Article |
| series | Acta Agriculturae Slovenica |
| spelling | doaj-art-68c8b73e40814e4bb4eb211be8547f742025-08-20T03:02:25ZengUniversity of Ljubljana Press (Založba Univerze v Ljubljani)Acta Agriculturae Slovenica1854-19411999-12-01742899810.14720/aas.1999.74.2.1590722300Development of cPCR technique for detection and enumeration of Prevotella bryantiiKatarina TEPŠIČ0Gorazd AVGUŠTIN1Univ. of Ljubljana, Biotechnical Fac., Zootechnical Dept., Groblje 3, SI-1230 Domžale, SloveniaUniv. of Ljubljana, Biotechnical Fac., Zootechnical Dept., Groblje 3, SI-1230 Domžale, SloveniaCompetitive PCR (cPCR) system for the detection and enumeration of Prevotella bryantii cells in rumen samples was developed. PBB14 primer, specific for P. bryantii was used as the reverse PCR primer and EUB 338 bacterial primer as the forward primer. An internal DNA control, containing primer specific sequences at 5’ and 3’ ends, but lacking 41 bp at the middle of the sequence, was constructed. C-PCR products were quantified by capillary electrophoresis and the results were calculated with regression equation (Reilly and Attwood, 1998). By coamplification of P.bryantii B14 genomic DNA extracted from known numbers of cells and internal control, standard curve was constructed which enables quantification of P.bryantii cells in the range of 2.15 x 103 to 4.3 x 104 cells.https://journals.uni-lj.si/aas/article/view/15907microbiologybacteriagram-negative bacteriaprevotella bryantiirumenanalytical chemistrycpcrcapillary electrophoresis |
| spellingShingle | Katarina TEPŠIČ Gorazd AVGUŠTIN Development of cPCR technique for detection and enumeration of Prevotella bryantii Acta Agriculturae Slovenica microbiology bacteria gram-negative bacteria prevotella bryantii rumen analytical chemistry cpcr capillary electrophoresis |
| title | Development of cPCR technique for detection and enumeration of Prevotella bryantii |
| title_full | Development of cPCR technique for detection and enumeration of Prevotella bryantii |
| title_fullStr | Development of cPCR technique for detection and enumeration of Prevotella bryantii |
| title_full_unstemmed | Development of cPCR technique for detection and enumeration of Prevotella bryantii |
| title_short | Development of cPCR technique for detection and enumeration of Prevotella bryantii |
| title_sort | development of cpcr technique for detection and enumeration of prevotella bryantii |
| topic | microbiology bacteria gram-negative bacteria prevotella bryantii rumen analytical chemistry cpcr capillary electrophoresis |
| url | https://journals.uni-lj.si/aas/article/view/15907 |
| work_keys_str_mv | AT katarinatepsic developmentofcpcrtechniquefordetectionandenumerationofprevotellabryantii AT gorazdavgustin developmentofcpcrtechniquefordetectionandenumerationofprevotellabryantii |