Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display
Differential display (DD) and the closely related RNA arbitrarily primed PCR (RAP-PCR) have become the molecular tools of choice for identifying and isolating differentially expressed genes in both eukaryotic and prokaryotic systems. However, one of the current drawbacks of both techniques is the hi...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2001-05-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/01305st04 |
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| _version_ | 1850152624158932992 |
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| author | Aaron C. Nagel James T. Fleming Gary S. Sayler Kenneth L. Beattie |
| author_facet | Aaron C. Nagel James T. Fleming Gary S. Sayler Kenneth L. Beattie |
| author_sort | Aaron C. Nagel |
| collection | DOAJ |
| description | Differential display (DD) and the closely related RNA arbitrarily primed PCR (RAP-PCR) have become the molecular tools of choice for identifying and isolating differentially expressed genes in both eukaryotic and prokaryotic systems. However, one of the current drawbacks of both techniques is the high number of false positives generated. In prokaryotic applications, the many false positives typically generated by DD are subsequently identified as rRNAs because of their greater abundance compared to mRNAs. To circumvent this problem, full-length 16S and 23S rDNA probes, derived from Pseudomonas putida G7 and Pseudomonas aeruginosa FRD1, respectively, were used as a prescreening approach to discriminate between those bands, which appear to be differentially expressed mRNAs, but in fact are rRNAs, following prokaryotic mRNA DD. |
| format | Article |
| id | doaj-art-e76e7cbbbe774faba08e8696fe7c6147 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2001-05-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-e76e7cbbbe774faba08e8696fe7c61472025-08-20T02:25:55ZengTaylor & Francis GroupBioTechniques0736-62051940-98182001-05-0130598899610.2144/01305st04Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential DisplayAaron C. Nagel0James T. Fleming1Gary S. Sayler2Kenneth L. Beattie31The University of Tennessee Knoxville, Knoxville, TN1The University of Tennessee Knoxville, Knoxville, TN1The University of Tennessee Knoxville, Knoxville, TN1The University of Tennessee Knoxville, Knoxville, TNDifferential display (DD) and the closely related RNA arbitrarily primed PCR (RAP-PCR) have become the molecular tools of choice for identifying and isolating differentially expressed genes in both eukaryotic and prokaryotic systems. However, one of the current drawbacks of both techniques is the high number of false positives generated. In prokaryotic applications, the many false positives typically generated by DD are subsequently identified as rRNAs because of their greater abundance compared to mRNAs. To circumvent this problem, full-length 16S and 23S rDNA probes, derived from Pseudomonas putida G7 and Pseudomonas aeruginosa FRD1, respectively, were used as a prescreening approach to discriminate between those bands, which appear to be differentially expressed mRNAs, but in fact are rRNAs, following prokaryotic mRNA DD.https://www.future-science.com/doi/10.2144/01305st04 |
| spellingShingle | Aaron C. Nagel James T. Fleming Gary S. Sayler Kenneth L. Beattie Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display BioTechniques |
| title | Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display |
| title_full | Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display |
| title_fullStr | Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display |
| title_full_unstemmed | Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display |
| title_short | Screening for Ribosomal-Based False Positives Following Prokaryotic mRNA Differential Display |
| title_sort | screening for ribosomal based false positives following prokaryotic mrna differential display |
| url | https://www.future-science.com/doi/10.2144/01305st04 |
| work_keys_str_mv | AT aaroncnagel screeningforribosomalbasedfalsepositivesfollowingprokaryoticmrnadifferentialdisplay AT jamestfleming screeningforribosomalbasedfalsepositivesfollowingprokaryoticmrnadifferentialdisplay AT garyssayler screeningforribosomalbasedfalsepositivesfollowingprokaryoticmrnadifferentialdisplay AT kennethlbeattie screeningforribosomalbasedfalsepositivesfollowingprokaryoticmrnadifferentialdisplay |