Differences in amplification efficiency of standard curves in quantitative real-time PCR assays and consequences for gene quantification in environmental samples
High and comparable efficiency values are the key for reliable quantification of target genes from environmental samples using real-time PCR. Therefore it was the aim of this study to investigate if PCR amplification efficiencies of plasmid DNA used for the calculation of standard curves (i) remain...
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| Veröffentlicht in: | Journal of microbiological methods 2010-09, Vol.82 (3), p.338-341 |
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| container_title | Journal of microbiological methods |
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| creator | Töwe, Stefanie Kleineidam, Kristina Schloter, Michael |
| description | High and comparable efficiency values are the key for reliable quantification of target genes from environmental samples using real-time PCR. Therefore it was the aim of this study to investigate if PCR amplification efficiencies of plasmid DNA used for the calculation of standard curves (i) remain constant along a logarithmic scale of dilutions and (ii) if these values are comparable to those of DNA extracted from environmental samples. It could be shown that comparable efficiency values within the standards cannot be achieved using log scale serial dilutions and a comparison of gene copy numbers from DNA extracted from environmental samples and standard DNA extracted from plasmids is only possible in a very small interval. |
| doi_str_mv | 10.1016/j.mimet.2010.07.005 |
| format | Article |
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Therefore it was the aim of this study to investigate if PCR amplification efficiencies of plasmid DNA used for the calculation of standard curves (i) remain constant along a logarithmic scale of dilutions and (ii) if these values are comparable to those of DNA extracted from environmental samples. 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All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c486t-c7dd488dc584f0a229dbd31fc6a671cfe1d6e8bb72f4cfa337c11ebd80f804bd3</citedby><cites>FETCH-LOGICAL-c486t-c7dd488dc584f0a229dbd31fc6a671cfe1d6e8bb72f4cfa337c11ebd80f804bd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.mimet.2010.07.005$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23217891$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20621132$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Töwe, Stefanie</creatorcontrib><creatorcontrib>Kleineidam, Kristina</creatorcontrib><creatorcontrib>Schloter, Michael</creatorcontrib><title>Differences in amplification efficiency of standard curves in quantitative real-time PCR assays and consequences for gene quantification in environmental samples</title><title>Journal of microbiological methods</title><addtitle>J Microbiol Methods</addtitle><description>High and comparable efficiency values are the key for reliable quantification of target genes from environmental samples using real-time PCR. Therefore it was the aim of this study to investigate if PCR amplification efficiencies of plasmid DNA used for the calculation of standard curves (i) remain constant along a logarithmic scale of dilutions and (ii) if these values are comparable to those of DNA extracted from environmental samples. It could be shown that comparable efficiency values within the standards cannot be achieved using log scale serial dilutions and a comparison of gene copy numbers from DNA extracted from environmental samples and standard DNA extracted from plasmids is only possible in a very small interval.</description><subject>Amplification efficiency</subject><subject>Bacteria - genetics</subject><subject>Bacteria - isolation & purification</subject><subject>Bacterial Proteins - genetics</subject><subject>Biological and medical sciences</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - isolation & purification</subject><subject>Environmental Microbiology</subject><subject>Functional genes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Dosage</subject><subject>LinRegPCR</subject><subject>Microbiology</subject><subject>Nitrogen cycle</subject><subject>Plasmids</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polymerase Chain Reaction - standards</subject><subject>Quantitative real-time PCR</subject><subject>Reference Standards</subject><subject>Standard curve</subject><issn>0167-7012</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc2KFDEUhYMoTjv6BIJkI66qzU9VJb1wIe0vDCii65BObiRNVaonN9XQj-Obmp5qx52uEnK_c-4hh5DnnK054_3r_XqMI5S1YPWFqTVj3QOy4lqJRstu85CsKqUaxbi4Ik8Q94zxTrb6MbkSrBecS7Eiv97FECBDcoA0JmrHwxBDdLbEKVEI9Rrr8ESnQLHY5G321M35uOC3s00llkofgWawQ1NqJvp1-41aRHtCWiXUTQnhdl6WhCnTn5Dgor3fVd0gHWOe0gip2IHiOQvgU_Io2AHh2eW8Jj8-vP--_dTcfPn4efv2pnGt7kvjlPet1t51ug3MCrHxOy95cL3tFXcBuO9B73ZKhNYFK6VynMPOaxY0ayt6TV4tvoc81axYzBjRwTDYBNOMRldjwYXk_yVVqze665SspFxIlyfEDMEcchxtPhnOzLlEszd3JZpziYYpU0usqhcX_3k3gr_X_GmtAi8vgEVnh5BtchH_clJwpTfnoG8WDuq_HSNkg3dtgo8ZXDF-iv8M8hucZcCn</recordid><startdate>20100901</startdate><enddate>20100901</enddate><creator>Töwe, Stefanie</creator><creator>Kleineidam, Kristina</creator><creator>Schloter, Michael</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20100901</creationdate><title>Differences in amplification efficiency of standard curves in quantitative real-time PCR assays and consequences for gene quantification in environmental samples</title><author>Töwe, Stefanie ; Kleineidam, Kristina ; Schloter, Michael</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c486t-c7dd488dc584f0a229dbd31fc6a671cfe1d6e8bb72f4cfa337c11ebd80f804bd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Amplification efficiency</topic><topic>Bacteria - genetics</topic><topic>Bacteria - isolation & purification</topic><topic>Bacterial Proteins - genetics</topic><topic>Biological and medical sciences</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - isolation & purification</topic><topic>Environmental Microbiology</topic><topic>Functional genes</topic><topic>Fundamental and applied biological sciences. 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| fulltext | fulltext |
| identifier | ISSN: 0167-7012 |
| ispartof | Journal of microbiological methods, 2010-09, Vol.82 (3), p.338-341 |
| issn | 0167-7012 1872-8359 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_858421231 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Amplification efficiency Bacteria - genetics Bacteria - isolation & purification Bacterial Proteins - genetics Biological and medical sciences DNA, Bacterial - genetics DNA, Bacterial - isolation & purification Environmental Microbiology Functional genes Fundamental and applied biological sciences. Psychology Gene Dosage LinRegPCR Microbiology Nitrogen cycle Plasmids Polymerase Chain Reaction - methods Polymerase Chain Reaction - standards Quantitative real-time PCR Reference Standards Standard curve |
| title | Differences in amplification efficiency of standard curves in quantitative real-time PCR assays and consequences for gene quantification in environmental samples |
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