Overview of the in situ production pipeline. (a-e) Examples illustrating each step of the high-throughput production of RNA in situ patterns. (a) Photograph of an agarose gel showing PCR products from a single 96-well plate. The vertical lines mark failed PCR reactions. (b) Nylon membrane spotted with 96 RNA in situ probes and stained to reveal the incorporation of digoxigenin. Missing or weak spots indicate a failed probe reaction. (c) Four wells of a 96-well plate, each containing about 200 embryos. (d) Low-resolution photograph of stained embryos. (e) High-resolution photograph of an individual embryo. All image and textual data are entered into a MySQL relational database and integrated with microarray data and previously published expression patterns.