Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4503

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Production of dsRNA for RNAi in Avian Embryos

Esther T. Stoeckli

This protocol was adapted from "RNAi in Avian Embryos," contributed by Esther T. Stoeckli, Chapter 14, in RNAi: A Guide to Gene Silencing (ed. Hannon). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.


INTRODUCTION

This protocol describes the production of double-stranded RNA (dsRNA) from fragments of cDNAs of candidate genes. The cDNA fragments must be cloned in plasmids with a flanking SP6 and T7 promoter (e.g., pSP72 or pCRII). The plasmid is linearized and sense and antisense RNAs are produced separately by in vitro transcription. After purification, the RNA strands are annealed to yield a dsRNA molecule suitable for RNAi in avian embryos.


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Related Protocol

Injection of dsRNA and Electroporation in Avian Embryos
Esther T. Stoeckli
CSH Protocols 2006: 4504. [Abstract] [Full Text]



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O. Mauti, T. Baeriswyl, and E. T. Stoeckli
Gene Silencing by Injection and Electroporation of dsRNA in Avian Embryos
CSH Protocols, December 1, 2008; 2008(13): pdb.prot5094 - pdb.prot5094.
[Abstract] [Full Text]