Format

Send to

Choose Destination
See comment in PubMed Commons below
Methods. 2001 Apr;23(4):359-72.

One-, two-, and three-color whole-mount in situ hybridization to Drosophila embryos.

Author information

1
Institut für Biologie I (Zoologie), Albert-Ludwigs-Universität Freiburg, Hauptstrasse 1, Freiburg im Breisgau, D-79104, Germany.

Abstract

This article contains detailed protocols for the localization of mRNA transcripts within whole Drosophila embryos. The procedures are based on the use of digoxigenin-, fluorescein-, and biotin-labeled antisense RNA probes for nonradioactive detection of transcripts. The labels are visualized in situ by differently colored water-insoluble precipitates using alkaline phosphatase- or beta-galactosidase-based immunoassays. First, a basic method is described that allows detection of transcript distribution(s) of one or more genes using the same color precipitate. Second, a sequential alkaline phosphatase detection method is presented that permits the visualization of two or three independent transcript patterns in multiple colors in the same embryo. Third, a shortened two-color in situ hybridization protocol is provided that employs a combination of beta-galactosidase and alkaline phosphatase colorimetric reactions for differential detection. The two-color in situ hybridization methods work equally well in Drosophila and zebrafish embryos and may therefore also be adaptable to other species.

PMID:
11316437
DOI:
10.1006/meth.2000.1148
[Indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Support Center