{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T21:14:03Z","timestamp":1791234843855,"version":"4.1.0"},"reference-count":39,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2011,1,1]],"date-time":"2011-01-01T00:00:00Z","timestamp":1293840000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2011,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Transcriptional analysis of microbial gene expression using relative quantitative real-time PCR (qRT-PCR) has been hampered by various technical problems. One such problem is the unavailability of an exogenous standard robust enough for use in a complex matrix like soil. To circumvent this technical issue, we made use of a recently developed artificial RNA (myIC) as an exogenous \u201cspike-in\u201d control. Nonsterile field soil was inoculated with various concentrations of the test bacterium\n            <jats:italic>Pseudomonas<\/jats:italic>\n            sp. strain LBUM300, ranging from 4.3- to 8.3-log bacterial cells per gram of soil. Total soil RNA was extracted at days 0, 7, and 14 postinoculation, and using two-step TaqMan assays,\n            <jats:italic>phlD<\/jats:italic>\n            (encoding the production of 2,4-diacetylphloroglucinol) and\n            <jats:italic>hcnC<\/jats:italic>\n            (encoding the production of hydrogen cyanide) gene expression was monitored. For relative quantification, a defined quantity of\n            <jats:italic>in vitro<\/jats:italic>\n            -synthesized myIC RNA was spiked during the RNA extraction procedure. Absolute qRT-PCR was also performed in parallel. Both the absolute and relative quantifications showed similar transcriptional trends. Overall, the transcriptional activity of\n            <jats:italic>phlD<\/jats:italic>\n            and\n            <jats:italic>hcnC<\/jats:italic>\n            changed over time and with respect to the bacterial concentrations used. Transcripts of the\n            <jats:italic>phlD<\/jats:italic>\n            and\n            <jats:italic>hcnC<\/jats:italic>\n            genes were detected for all five bacterial concentrations, but the\n            <jats:italic>phlD<\/jats:italic>\n            transcript copy numbers detected were lower than those detected for\n            <jats:italic>hcnC<\/jats:italic>\n            , regardless of the initial bacterial concentration or sampling date. For quantifying a low number of transcripts, the relative method was more reliable than the absolute method. This study demonstrates for the first time the use of a relative quantification approach to quantifying microbial gene transcripts from field soil using an exogenous spike-in control.\n          <\/jats:p>","DOI":"10.1128\/aem.01387-10","type":"journal-article","created":{"date-parts":[[2010,11,13]],"date-time":"2010-11-13T05:59:41Z","timestamp":1289627981000},"page":"41-47","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":31,"title":["Relative and Absolute Quantitative Real-Time PCR-Based Quantifications of\n            <i>hcnC<\/i>\n            and\n            <i>phlD<\/i>\n            Gene Transcripts in Natural Soil Spiked with\n            <i>Pseudomonas<\/i>\n            sp. Strain LBUM300"],"prefix":"10.1128","volume":"77","author":[{"given":"Nadine J.","family":"DeCoste","sequence":"first","affiliation":[{"name":"Universite\u0301 de Moncton, Department of Biology, 18 Antonine-Maillet, Moncton, NB E1A 3E9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vijay J.","family":"Gadkar","sequence":"additional","affiliation":[{"name":"Universite\u0301 de Moncton, Department of Biology, 18 Antonine-Maillet, Moncton, NB E1A 3E9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Filion","sequence":"additional","affiliation":[{"name":"Universite\u0301 de Moncton, Department of Biology, 18 Antonine-Maillet, Moncton, NB E1A 3E9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","unstructured":"The Canadian System of Soil Classification 3rd ed. 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