Preparation of dsRNA for microinjection experiments in mouse

. 2009 Jan ; 2009 (1) : pdb.prot5131.

Jazyk angličtina Země Spojené státy americké Médium print

Typ dokumentu časopisecké články

Perzistentní odkaz   https://www.medvik.cz/link/pmid20147026
Odkazy

PubMed 20147026
DOI 10.1101/pdb.prot5131
PII: 2009/1/pdb.prot5131
Knihovny.cz E-zdroje

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.

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