Preparation of dsRNA for microinjection experiments in mouse
Language English Country United States Media print
Document type Journal Article
PubMed
20147026
DOI
10.1101/pdb.prot5131
PII: 2009/1/pdb.prot5131
Knihovny.cz E-resources
- MeSH
- RNA, Double-Stranded genetics MeSH
- Transcription, Genetic MeSH
- Genetic Techniques * MeSH
- Microinjections methods MeSH
- Models, Genetic MeSH
- Molecular Biology methods MeSH
- Molecular Sequence Data MeSH
- Mice MeSH
- Oocytes metabolism MeSH
- Plasmids metabolism MeSH
- Polymerase Chain Reaction methods MeSH
- RNA Interference MeSH
- RNA metabolism MeSH
- Base Sequence MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- RNA, Double-Stranded MeSH
- RNA MeSH
RNA interference (RNAi) is a suitable method for sequence-specific post-transcriptional gene silencing for a number of model systems. The protocol presented here is designed for the preparation of microgram amounts of double-stranded RNA (dsRNA) for microinjection experiments in mouse oocytes and early embryos. Briefly, dsRNA is produced after annealing sense and antisense RNA strands, or spontaneously during in vitro transcription of an inverted repeat. We usually include RNase T1 treatment of the annealed RNA prior to the purification step in order to remove unannealed single-stranded RNA (ssRNA), which can interfere with the quantification of dsRNA in a nondenaturing agarose gel.
References provided by Crossref.org
Age-related differences in the translational landscape of mammalian oocytes
