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The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in a partnership with Sigma-Aldrich. A list of products is available to use in each protocol.
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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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refer to the detailed product description on the usage of specific products of interest.
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