Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5131

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Preparation of dsRNA for Microinjection Experiments in Mouse

Petr Svoboda

Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague 4, Czech Republic

Corresponding author (svobodap{at}img.cas.cz)


INTRODUCTION

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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This article has been cited by other articles:


Home page
 CSH ProtocolsHome page
P. Svoboda
Choosing the Sequence of dsRNA for RNAi in Mouse
CSH Protocols, January 1, 2009; 2009(1): pdb.ip63 - pdb.ip63.
[Abstract] [Full Text]

Home page
 CSH ProtocolsHome page
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CSH Protocols, January 1, 2009; 2009(1): pdb.ip64 - pdb.ip64.
[Abstract] [Full Text]

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 CSH ProtocolsHome page
P. Svoboda and P. Stein
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CSH Protocols, January 1, 2009; 2009(1): pdb.top56 - pdb.top56.
[Abstract] [Full Text]

Home page
 CSH ProtocolsHome page
P. Stein
Microinjection of dsRNA into Fully-Grown Mouse Oocytes
CSH Protocols, January 1, 2009; 2009(1): pdb.prot5132 - pdb.prot5132.
[Abstract] [Full Text]

Home page
 CSH ProtocolsHome page
P. Stein
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CSH Protocols, January 1, 2009; 2009(1): pdb.prot5133 - pdb.prot5133.
[Abstract] [Full Text]