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Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success

(BIRC5) or Silencer Negative Control #1 in quadruplicate. Forty-eight hours later, two sets of samples were stained with DAPI and analyzed by fluorescence microscopy. Total RNA was isolated from the other two sample sets using RNAqueous MAG-96 and then converted to cDNA. Target cDNA levels were analyzed in duplicate using a TaqMan Gene Expression Assay for survivin (Hs00153353_m1). Data shown here represent duplicate assays from single transfections. Survivin mRNA levels were reduced 80% in the survivin siRNA treated cells relative to control siRNA treated cells. Knockdown of survivin induced morphological changes to cell nuclei. The insets show representative cells from the survivin and control siRNA treated samples.


Summary
Thanks to optimized siRNA and real-time assay design using sophisticated bioinformatics and design algorithms, performing gene silencing experiments is now easier than ever. Pre-designed siRNA and qRT-PCR assay combinations are linked through Ambion's website. The siRNA Database provides information about Ambion's Silencer Pre-designed siRNAs targeting almost all human, mouse, and rat genes in NCBI's RefSeq database. The siRNA database is also a conduit to Silencer Validated siRNAs, which have been functionally proven to reduce target mRNA levels by >70%. To find Silencer siRNAs and TaqMan Gene Expression Assays to your gene of interest, visit www.ambion.com/siRNA.


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