Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
To the authors’ knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studi...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-07-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/8/8/1099 |
| _version_ | 1797561604631429120 |
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| author | Lucilla Iacumin Francesca Cecchini Marco Vendrame Giuseppe Comi |
| author_facet | Lucilla Iacumin Francesca Cecchini Marco Vendrame Giuseppe Comi |
| author_sort | Lucilla Iacumin |
| collection | DOAJ |
| description | To the authors’ knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studies was tested. Our results indicated that ePCR results in uniform amplification of several DNA molecules, overcoming the major limitations of conventional PCR, such as preferential amplification and DNA concentration dependence. Moreover, ePCR-DGGE resulted in higher sensitivity when compared to conventional PCR-DGGE methods used for studying microbial populations in a complex matrix. In fact, compared to conventional PCR, the DGGE profiles of ePCR products permitted the detection of a higher number of the species that were present in the tested sample. |
| first_indexed | 2024-03-10T18:17:17Z |
| format | Article |
| id | doaj.art-c9abeb52ce9f49cdb41be4c436a91d03 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T18:17:17Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-c9abeb52ce9f49cdb41be4c436a91d032023-11-20T07:38:58ZengMDPI AGMicroorganisms2076-26072020-07-0188109910.3390/microorganisms8081099Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population StudiesLucilla Iacumin0Francesca Cecchini1Marco Vendrame2Giuseppe Comi3Department of Agricultural, Food, Environmental and Animal Science, University of Udine, via Sondrio 2/A, 33100 Udine, ItalyDepartment of Agricultural, Food, Environmental and Animal Science, University of Udine, via Sondrio 2/A, 33100 Udine, ItalyDepartment of Agricultural, Food, Environmental and Animal Science, University of Udine, via Sondrio 2/A, 33100 Udine, ItalyDepartment of Agricultural, Food, Environmental and Animal Science, University of Udine, via Sondrio 2/A, 33100 Udine, ItalyTo the authors’ knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studies was tested. Our results indicated that ePCR results in uniform amplification of several DNA molecules, overcoming the major limitations of conventional PCR, such as preferential amplification and DNA concentration dependence. Moreover, ePCR-DGGE resulted in higher sensitivity when compared to conventional PCR-DGGE methods used for studying microbial populations in a complex matrix. In fact, compared to conventional PCR, the DGGE profiles of ePCR products permitted the detection of a higher number of the species that were present in the tested sample.https://www.mdpi.com/2076-2607/8/8/1099emulsion-PCRDGGEdirect molecular methodsbacterial ecology |
| spellingShingle | Lucilla Iacumin Francesca Cecchini Marco Vendrame Giuseppe Comi Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies Microorganisms emulsion-PCR DGGE direct molecular methods bacterial ecology |
| title | Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies |
| title_full | Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies |
| title_fullStr | Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies |
| title_full_unstemmed | Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies |
| title_short | Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies |
| title_sort | emulsion pcr epcr as a tool to improve the power of dgge analysis for microbial population studies |
| topic | emulsion-PCR DGGE direct molecular methods bacterial ecology |
| url | https://www.mdpi.com/2076-2607/8/8/1099 |
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