( h RNA and protein from your RNAi experi...)RNAi: Get the Whole Story

Figure 3. Timecourse of PCNA siRNA Effects on PCNA mRNA and Protein Levels. HeLa cells were plated at 30,000 cells/well into a 24 well culture plate. The cells were transfected with an siRNA against PCNA or with the Silencer Negative Control #1 (scrambled) at a final concentration of 100 nM using siPORT Lipid. Duplicate samples were harvested at 72 hours and each day for 5 days. The PARIS Kit was used to isolate both RNA and protein for either real-time RT-PCR (Panel A) or Western blot analysis (Panel B). RNA was reverse transcribed using the RETROscript Kit; and target cDNA levels were measured by real-time PCR using SYBR Green assays. Input cDNA in the different samples was normalized using real-time data for 18S rRNA. The bar graphs represent an average of three data points. Data are shown relative to mRNA levels of cells transfected with scrambled control siRNA.

Tools to Assist You in Your RNAi Research
Ambion offers products for analysis of both mRNA and protein from the same sample; here we demonstrate the versatility of the PARIS Kit for this application. Other Ambion products used in this study include Silencer Validated siRNAs to Stat-1, p53, and PCNA; and Silencer GAPDH control siRNAs for eliciting RNAi. Also used w
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