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  Large-scale expression screening by automated whole-mount in situ hybridization

Quiring, R., Wittbrodt, B., Henrich, T., Ramialison, M., Burgtorf, C., Lehrach, H., et al. (2004). Large-scale expression screening by automated whole-mount in situ hybridization. Mechanisms of Development, 121(7-8), 971-976. doi:10.1016/j.mod.2004.03.031.

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Genre: Journal Article
Alternative Title : Mech Dev

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 Creators:
Quiring, Rebecca, Author
Wittbrodt, Beate, Author
Henrich, Thorsten, Author
Ramialison, Mirana, Author
Burgtorf, Carola1, Author
Lehrach, Hans2, Author           
Wittbrodt, Joachim, Author
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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Free keywords: Oryzias latipes; Partial cDNA sequencing; Expressed Sequence Tags; Screening; Automated; Whole-mount in situ hybridization; Gene expression; High-throughput; Database; Expression profile; Embryogenesis; Organogenesis
 Abstract: Gene expression profiling is an important component of functional genomics. We present a time and cost efficient high-throughput whole-mount in situ technique to perform a large-scale gene expression analysis in medaka fish (Oryzias latipes) embryos. Medaka is a model system ideally suited for the study of molecular genetics of vertebrate development. Random cDNA clones from an arrayed stage 20 medaka plasmid library were analyzed by whole-mount in situ hybridization on embryos of three representative stages of medaka development. cDNA inserts were colony PCR amplified in a 384-format. The PCR products were used to generate over 2000 antisense RNA digoxigenin probes in a high-throughput process. Whole-mount in situ hybridization was carried out in a robot and a broad range of expression patterns was observed. Partial cDNA sequences and expression patterns were documented with BLAST results, cluster analysis, images and descriptions, respectively; collectively this information was entered into a web-based database, ‘MEPD’ (http://www.embl-heidelberg.de/mepd/), that is publicly accessible.

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Language(s): eng - English
 Dates: 2004-07
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: Mechanisms of Development
  Alternative Title : Mech Dev
Source Genre: Journal
 Creator(s):
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Publ. Info: -
Pages: - Volume / Issue: 121 (7-8) Sequence Number: - Start / End Page: 971 - 976 Identifier: ISSN: 0925-4773