| |  Online-Ressource |
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| Verfasst von: | Ghafoory, Shahrouz [VerfasserIn]  |
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| | Breitkopf-Heinlein, Katja [VerfasserIn] |
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| | Li, Qi [VerfasserIn] |
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| | Dzieran, Johanna [VerfasserIn] |
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| | Scholl, Catharina [VerfasserIn] |
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| | Dooley, Steven [VerfasserIn] |
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| | Wölfl, Stefan [VerfasserIn] |
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| Titel: | A fast and efficient polymerase chain reaction-based method for the preparation of in situ hybridization probes |
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| Verf.angabe: | Shahrouz Ghafoory, Katja Breitkopf‐Heinlein, Qi Li, Johanna Dzieran, Catharina Scholl, Steven Dooley & Stefan Wölfl |
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| E-Jahr: | 2012 |
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| Jahr: | 28 March 2012 |
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| Umfang: | 8 S. |
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| Fussnoten: | Gesehen am 07.06.2018 |
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| Titel Quelle: | Enthalten in: Histopathology |
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| Ort Quelle: | Oxford [u.a.] : Wiley-Blackwell, 1977 |
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| Jahr Quelle: | 2012 |
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| Band/Heft Quelle: | 61(2012), 2, Seite 306-313 |
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| ISSN Quelle: | 1365-2559 |
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| Abstract: | Ghafoory S, Breitkopf-Heinlein K, Li Q, Dzieran J, Scholl C, Dooley S & Wölfl S (2012) Histopathology 61, 306-313 A fast and efficient polymerase chain reaction-based method for the preparation of in situ hybridization probes Aims: In situ hybridization (ISH) is the method of choice for analysis of the local distribution of gene expression in tissue samples at the cellular level. In this study we present a rapid and efficient protocol for the generation of labelled cRNA probes. Methods and results: The protocol is based on the preparation of DNA in vitro transcription templates using polymerase chain reaction (PCR), using primers that include RNA polymerase promoter sequences and size-based purification of PCR fragments containing the target gene-specific cDNA and promoter elements for T7 and SP6 RNA polymerase. The optimized purification protocols ensure high transcription efficiency and target specificity of the labelled cRNA. The cRNA hybridization probes obtained are compatible with established in situ hybridization protocols. Conclusions: Purified PCR fragment-based in vitro transcription enables preparation of in situ hybridization probes which allow the rapid detection of gene expression distribution in tissue slices from any gene of interest. |
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| DOI: | doi:10.1111/j.1365-2559.2012.04237.x |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: http://dx.doi.org/10.1111/j.1365-2559.2012.04237.x |
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| | Volltext: https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1365-2559.2012.04237.x |
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| | DOI: https://doi.org/10.1111/j.1365-2559.2012.04237.x |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | albumin |
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| | alpha-fetoprotein |
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| | in situ hybridization |
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| | polymerase chain reaction |
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| | primer |
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| K10plus-PPN: | 1576131718 |
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| Verknüpfungen: | → Zeitschrift |
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¬A¬ fast and efficient polymerase chain reaction-based method for the preparation of in situ hybridization probes / Ghafoory, Shahrouz [VerfasserIn]; 28 March 2012 (Online-Ressource)
