Functional Genomics and RNAi

Production of dsRNA for RNAi

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Selected Protocol from "RNAi, A Guide to Gene Silencing", Esther T. Stoeckli.
RNAi in Avian Embryos, Esther T. Stoeckli (Chapter 14)

Production of dsRNA for RNAi in Avian Embryos

ABSTRACT
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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Products Available for this Protocol

Protocol Material Description Product #  Product Name Add to Cart
Buffers, Solutions, and Reagents      
Acidic phenol:chloroform (phenol:chloroform:isoamyl alcohol 25:24:1) 77617 Phenol – chloroform – isoamyl alcohol mixture, for molecular biology, 25:24:1
Ammonium acetate A1542 Ammonium acetate, for molecular biology
Chloroform:isoamyl alcohol (24:1) C0549 Chloroform:Isoamyl alcohol 24:1
EDTA E7889 Ethylenediaminetetraacetic acid disodium salt solution
DNase I S3014 DNase I, Amplification Grade
Ethanol P9541 Ethanol, for molecular biology, 190 proof
Phosphate-buffered saline (PBS) P9541 Phosphate buffered saline, pH 7.4, Biotechnology Performance Certified

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