BLOCK-iT™ Complete Dicer RNAi Kit
from
Invitrogen
Description
The BLOCK-iT™ Dicer RNAi Transfection Kit provides a quick, easy alternative to using synthetic short RNA oligonucleotides for RNAi experiments. Effective blocking of gene expression in mammalian cells requires the use of siRNA (1). dsRNA transcribed in vitro can be cleaved into siRNA by the addition of purified, recombinant Dicer protein in vitro (2-5). Theres no need to know the best sequence to get the desired RNAi effect and no time spent on siRNA primer design, making this approach ideal for initial screening to determine RNAi results. The BLOCK-iT™ Dicer RNAi Transfection Kit provides everything needed to obtain and transfect a pool of siRNA specific to your gene, including:
- A high-quality preparation of Dicer enzyme that results in high yields of a 21-23 nucleotide d-siRNA pool to produce the desired RNAi effect
- An siRNA purification module specifically developed to isolate pure siRNA from the Dicer reaction
- Lipofectamine™ 2000 Transfection Reagent for proven high-efficiency transfection-essential for robust gene knockdown-in a wide variety of mammalian cells
- A BLOCK-iT™ RNAi TOPO® Transcription Kit for rapid, high-yield synthesis of long double-stranded RNA (BLOCK-iT™ Complete Dicer RNAi Kit only)
It is important to use high-purity siRNA in order to achieve the desired RNAi effect. The BLOCK-iT™ Complete Dicer RNAi Kit combines all the necessary reagents in a single system to obtain high-purity siRNA for specific and effective knockdown of gene expression (Figure 1). The BLOCK-iT™ Dicer Kits provide an efficient and cost-effective way to quickly screen as many as five genes in a mammalian model system.
How BLOCK-iT™ Dicer Transfection WorksLong dsRNA transcribed
in vitro with the BLOCK-iT™ RNAi TOPO¤ Transcription Kit, or by another method, is used as a substrate for the BLOCK-iT™ Dicer reaction. Following addition of the BLOCK-iT™ Dicer Enzyme, the long dsRNA will be cleaved into 21-23 nucleotide siRNA fragments (Figure 2). The processing of long dsRNA by Dicer generates siRNA throughout the initial substrate. Each reaction results in a pool of siRNA, a number of which should be highly effective siRNA to the target sequence. More highly effective siRNA duplexes are likely to be present in this pool of siRNA than if a single siRNA target site is selected, giving the pool itself a high activity in transient RNAi responses. Following transfection, the specific reduction of targeted gene expression is assayed (Figure 1).