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Citation
Weiszmann, R., Hammonds, A.S., Celniker, S.E. (2009). Determination of gene expression patterns using high-throughput RNA in situ hybridization to whole-mount Drosophila embryos.  Nat. Protoc. 4(5): 605--618.
FlyBase ID
FBrf0207749
Publication Type
Research paper
Abstract

We describe a high-throughput protocol for RNA in situ hybridization (ISH) to Drosophila embryos in a 96-well format. cDNA or genomic DNA templates are amplified by PCR and then digoxigenin-labeled ribonucleotides are incorporated into antisense RNA probes by in vitro transcription. The quality of each probe is evaluated before ISH using a RNA probe quantification (dot blot) assay. RNA probes are hybridized to fixed, mixed-staged Drosophila embryos in 96-well plates. The resulting stained embryos can be examined and photographed immediately or stored at 4 degrees C for later analysis. Starting with fixed, staged embryos, the protocol takes 6 d from probe template production through hybridization. Preparation of fixed embryos requires a minimum of 2 weeks to collect embryos representing all stages. The method has been used to determine the expression patterns of over 6,000 genes throughout embryogenesis.

PubMed ID
PubMed Central ID
PMC2780369 (PMC) (EuropePMC)
Related Publication(s)
Website

BDGP insitu homepage
Fisher et al., 2012, BDGP insitu homepage [FBrf0219073]

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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    Nat. Protoc.
    Title
    Nature Protocols
    Publication Year
    2006--
    ISBN/ISSN
    1754-2189 1750-2799
    Data From Reference