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Spatial expression profiles in the Xenopus laevis oocytes measured with qPCR tomography
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SYSNO ASEP 0346483 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Spatial expression profiles in the Xenopus laevis oocytes measured with qPCR tomography Author(s) Šindelka, R. (GB)
Šídová, Monika (BTO-N) RID
Švec, David (BTO-N) RID
Kubista, Mikael (BTO-N) RIDSource Title Methods. - : Elsevier - ISSN 1046-2023
Roč. 51, - (2010), s. 87-91Number of pages 5 s. Language eng - English Country US - United States Keywords Xenopus ; Oocyte ; qPCR tomography Subject RIV EB - Genetics ; Molecular Biology R&D Projects IAA500970904 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) GA301/09/1752 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z50520701 - BTO-N (2007-2013) UT WOS 000277953300013 Annotation qPCR tomography was developed to study mRNA localization in complex biological samples that are embedded and cryo-sectioned. After total RNA extraction and reverse transcription, the spatial profiles of mRNAs and other functional RNAs were determined by qPCR. The Xenopus laevis oocyte was selected as model, because of its large size (more than 1 mm) and large amount of total RNA ( 5 μg). Fifteen sections along the animal–vegetal axis were cut and prepared for quantification of 31 RNA targets using the high-throughput real-time RT-PCR (qPCR) BioMark™ platform. mRNAs were found to have two localization patterns, animal/central or vegetal. Because of the high resolution in sectioning, it was possible to distinguish two subgroups of the vegetal gene patterns: germ plasm determinant pattern and profile of other vegetal genes Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2011
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