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RNAi: A How To for New Users

purpose. Ambion's siRNA database provides links to individual assays matched to gene specific Silencer Pre-designed and Validated siRNAs, which makes finding both siRNAs and real-time PCR assays to your gene of interest extremely fast and easy. Additional time can be saved with the Cells-to-Signal Kit. Typically RNA isolation is performed prior to qRT-PCR. The Cells-to-Signal Kit allows you to perform qRT-PCR directly in lysates without RNA isolation. The Cells-to-Signal procedure is compatible with TaqMan Gene Expression Assays.

Although it is necessary to monitor mRNA levels to validate siRNAs, most researchers also want to determine the extent of knockdown at the protein level. Ambions PARIS Kit (Protein And RNA Isolation System) provides a simple method for isolating total RNA and protein from the same sample. RNA isolated with the kit can be used for RT-PCR, array analysis, Northern blotting, or other analytical techniques, and the protein lysate is compatible with Western blotting. Since native protein is recovered in most cases, enzymatic assays can also be performed.

The most careful of researchers correlate siRNA, target mRNA, and target protein levels. Isolat
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