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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 |
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| Acidic phenol:chloroform (phenol:chloroform:isoamyl alcohol 25:24:1) |
77617 |
Phenol – chloroform – isoamyl alcohol mixture, for molecular biology, 25:24:1 |
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| Ammonium acetate |
A1542 |
Ammonium acetate, for molecular biology |
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| Chloroform:isoamyl alcohol (24:1) |
C0549 |
Chloroform:Isoamyl alcohol 24:1 |
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| EDTA |
E7889 |
Ethylenediaminetetraacetic acid disodium salt solution |
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| DNase I |
S3014 |
DNase I, Amplification Grade |
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| Ethanol |
P9541 |
Ethanol, for molecular biology, 190 proof |
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| Phosphate-buffered saline (PBS) |
P9541 |
Phosphate buffered saline, pH 7.4, Biotechnology Performance Certified |
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Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.
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