Thermo Fisher, V47020, pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit

pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit Using the pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit, you can:
•Assess the potency of any RNAi knockdown reagent using a fast, sensitive, and cost-effective β-gal reporter assay
•Identify the best knockdown reagent for your RNAi experiments without knowing protein function, western blotting, or qRT-PCRHow it Works:The BLOCK-iT™ RNAi Target Screening system method uses a β-galactosidase activity assay to measure the knockdown ability of an RNAi reagent (Stealth RNAi™ siRNA, siRNA, dicer pools, or shRNA-containing plasmid). To identify the most effective knockdown reagent for your target, simply clone your target gene into the pSCREEN-iT™/lacZ-DEST Gateway™ vector. Co-transfect the resulting construct with the knockdown reagent being tested. Expressed genes will be fused to β-galactosidase, enabling you to use β-galactosidase levels to measure the amount of RNAi-induced degradation of the target gene (Figure 1). The more effective any specific knockdown reagent is at mediating RNAi, the more degradation of the fusion mRNA will occur, lowering β-gal activity (Figure 2).
The BLOCK-iT™ RNAi Target Screening System includes the pSCREEN-iT™/lacZ-DEST Gateway™ vector for generating a construct containing alacZ/gene of interest fusion. This vector features:
•attR sites immediately following thelacZ ORF for fast and efficient recombination with anyattL-flanked Gateway™ entry vector to generate thelacZ/target gene fusions
•The CMV promoter for strong expression of thelacZ/target gene fusion, leading to increased assay sensitivity and better discrimination between knockdown reagents of varied effectiveness
•Positional cloning to fuse a gene or fragment to the 3´ end of thelacZ gene so that β-galactosidase will be expressed even if the target fragment being tested contains stop codonsIn addition, the pSCREEN-iT™/lacZ-DEST vector is fully compatible with the Ultimate™ ORF Clone Collection. This makes a wide selection of potential target genes readily available for generatinglacZ fusions.
For Use With (Application): RNAi Target Screening
Format: Kit
Product Type: β-Galactosidase Reporter Gene Assay System
Quantity: 20 Reactions
Reporter Gene: Beta-Gal (lacZ)
Sample Type: Synthetic siRNA, RNAi Plasmids (shRNA, miR)
Product Line: BLOCK-iT™, Gateway™
Promoter: CMV
Unit Size: 20 reactions