In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea.

ABSTRACT:

Background

The freshwater planarian Schmidtea mediterranea has emerged as a powerful model for studies of regenerative, stem cell, and germ cell biology. Whole-mount in situ hybridization (WISH) and whole-mount fluorescent in situ hybridization (FISH) are critical methods for determining gene expression patterns in planarians. While expression patterns for a number of genes have been elucidated using established protocols, determining the expression patterns for particularly low-abundance transcripts remains a challenge.

Results

We show here that a short bleaching step in formamide dramatically enhances signal intensity of WISH and FISH. To further improve signal sensitivity we optimized blocking conditions for multiple anti-hapten antibodies, developed a copper sulfate quen

SUBMITTER: King RS 

PROVIDER: S-EPMC3610298 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature