Cost Effective High Potency SiRNAs

Cost Effective High Potency siRNAs
Silencer™ siRNA Construction Kit

• Make 15 siRNAs for the cost of chemically synthesizing just 1 siRNA

• Hundreds of transfections per reaction

• Fully licensed for siRNA research

• 10X­20X the potency of chemically synthesized siRNAs

In order to develop the most effective siRNA to your gene target, several siRNA molecules need to be designed and tested. This could require chemical synthesis and purification of 6-10 RNA oligonucleotides at an estimated cost of $1500 per gene. Ambion has developed a protocol for siRNA synthesis and purification based on in vitro transcription -- the Silencer™ siRNA Construction Kit. The kit produces transfection ready siRNA at a fraction of the cost of chemical synthesis, enabling screening of more sequence within your gene for identification of the most potent siRNA.

Construction of an siRNA requires the synthesis of two inexpensive, desalted DNA oligonucleotides. These DNA oligonucleotides include an 8 base sequence complementary to the 5' end of the T7 promoter primer included in the Silencer siRNA Construction Kit. Each gene specific oligonucleotide is used to generate a double-stranded template for use in the in vitro transcription step. The two transcription reaction products, representing sense and antisense strands, are hybridized to each other and processed to yield transfection ready siRNA. The entire procedure requires less than 2 days, is easily scalable (15 siRNA molecules can be synthesized as easily as a single siRNA), and produces enough siRNA for hundreds of transfections.

The Silencer siRNA Construction Kit includes all necessary reagents to generate 15 double-stranded siRNAs, ready for transfection. Also included are a detailed Instruction Manual and a GAPDH control template. The GAPDH specific siRNA made in the control reaction is functional in both human and mouse cells and can be used for testing and troubleshooting transfection reagents and protocols.