In Vitro Transcribed (IVT) siRNA

A Complete IVT siRNA Synthesis Solution
The Silencer siRNA Construction Kit includes all necessary reagents to generate and purify up to 15 double-stranded siRNAs, ready for transfection. Also included are a detailed Instruction Manual and a GAPDH siRNA control template. The GAPDH-specific siRNA made in the control reaction can be used for optimizing transfection protocols.

How the Kit Works
The in vitro transcription method included with the Silencer siRNA Construction Kit uses T7 RNA polymerase to generate individual strands of the siRNA.

1. Templates for the reactions are produced from two inexpensive DNA oligonucleotides encoding the desired siRNA strands. These oligonucleotides are designed to include an 8 base sequence complementary to the 3' end of the T7 promoter primer included in the kit.
2. The oligonucleotides are each annealed to the T7 promoter primer, and a fill-in reaction with Klenow fragment generates a double-stranded template ready for use in the in vitro transcription reaction.
3. After transcription, the reactions are combined to permit annealing of the two siRNA strands.
4. The siRNA preparation is then treated with DNase (to remove template) and RNase (to polish the ends of the double-stranded RNA), and then column purified. The entire procedure requires very little hands-on time and can be completed in less than 24 hours.