Protocol

Transcript In Situ Hybridization of Whole-Mount Embryos for Phenotype Analysis of RNAi-Treated Drosophila

This protocol was adapted from "RNAi in Drosophila" contributed by Richard W. Carthew, Chapter 17, in RNAi, A Guide to Gene Silencing (ed. Hannon). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 100 words of the full text of this article appear below.

INTRODUCTION

This protocol fixes and prepares embryos for in situ hybridization to visualize transcript expression patterns. It is a modification of the method developed by Tautz and Pfeifle for whole-mount in situ analysis of embryos. Use of the standard hybridization protocol on RNAi-treated embryos results in high background staining, which makes visualization of transcript expression patterns practically impossible. The following modifications eliminate this problem and allow visualization of transcript expression after RNAi injections.

MATERIALS

Reagents

Embryos (from Microinjection of dsRNA into Drosophila Embryos and Delivery of dsRNA into Drosophila Embryos by a Gene Gun)

Fixative A

Fixative B

Formamide/TE

Glycine block

Halocarbon . . . [Full Text of this Article]

Equipment

METHOD