Format

Send to

Choose Destination
Methods Mol Biol. 2017;1605:121-131. doi: 10.1007/978-1-4939-6988-3_8.

Purification of Zygotically Transcribed RNA through Metabolic Labeling of Early Zebrafish Embryos.

Author information

1
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307, Dresden, Germany. Patricia.Heyn@igmm.ed.ac.uk.
2
MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh, EH4 2XU, UK. Patricia.Heyn@igmm.ed.ac.uk.
3
Molecular Biophysics & Biochemistry, Yale University, 333 Cedar St, New Haven, CT, 06520, USA. karla.neugebauer@yale.edu.

Abstract

Early embryonic development in all known metazoans is characterized by a transcriptionally silent phase, during which development is under control of maternally loaded protein and RNA. The zygotic genome becomes transcriptionally active after a series of rapid reductive cleavage divisions. In this chapter, we present a method to metabolically label, purify, and analyze newly transcribed RNAs in early zebrafish embryos. We previously used this method, which is adaptable to other embryos and systems, to determine the onset of zygotic transcription activation and identify the first zygotic transcripts.

KEYWORDS:

4-sUTP; Biotinylation; RNA; Transcription; Zebrafish; Zygotic genome activation

PMID:
28456961
DOI:
10.1007/978-1-4939-6988-3_8
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center