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.
Original language | English |
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Pages (from-to) | 971-976 |
Number of pages | 6 |
Journal | Mechanisms of Development |
Volume | 121 |
Issue number | 7-8 |
DOIs | |
Publication status | Published - Jul 2004 |
Externally published | Yes |
Keywords
- Automated
- Database
- Embryogenesis
- Expressed Sequence Tags
- Expression profile
- Gene expression
- High-throughput
- Organogenesis
- Oryzias latipes
- Partial cDNA sequencing
- Screening
- Whole-mount in situ hybridization