Abstract
INTRODUCTIONThis in situ hybridization (ISH) protocol describes a simplified method using a digoxigenin-labeled antisense RNA probe on whole Xenopus embryos, suitable for both X. laevis and X. tropicalis. The protocol includes fixation, β-galactosidase staining (when lineage tracing is needed), and storage of the embryos prior to ISH. This method shortens the steps before hybridization, which limits RNA degradation in the sample, and preserves superficial structures. Hence, it is particularly suited for the analysis of ectoderm, neural, and mesodermal structures from blastula to early tadpole stages. Additional permeabilization steps are included to process later tadpole stages.
Publisher
Cold Spring Harbor Laboratory
Subject
General Biochemistry, Genetics and Molecular Biology
Reference3 articles.
1. In situ hybridization: An improved whole-mount method for Xenopus embryos;Harland;Methods Cell Biol.,1991
2. Nieuwkoop P.D. Faber J. (1994) Normal table of Xenopus laevis (Daudin) (Garland, New York).
3. Sive H.L. Grainger R.M. Harland R.M. (2000) Early development of Xenopus laevis: A laboratory manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.)
Cited by
34 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献