Invitrogen™ pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit

Invitrogen™ pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit

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 

Features

• 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-PCR

Contents

• The pSCREEN-iT™ /lacZ-DEST Gateway™ Vector Kit contains 6 µg of vector and 10 µg of pSCREEN-iT™/lacZ-GW/CDK2 control vector, the positive control Stealth™RNAi lacZ, a negative control Scrambled Stealth RNAi™ siRNA, and 1X annealing/dilution buffer. Store at -20°C. 
• The BLOCK-iT™ RNAi Target Screening Kit contains the pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit, 250 µl of Lipofectamine™ 2000 Reagent, and the FluoReporter™ lacZ/Galactosidase Quantification Kit. Store the Lipofectamine™ 2000 Reagent at +4°C. Store the FluoReporter™ Kit at -20°C protected from light. 
• The BLOCK-iT™ RNAi Target Screening System contains the BLOCK-iT™ RNAi Target Screening Kit, LR Clonase™ enzyme mix, and pCR ¤ 8/GW/TOPO ¤ TA Cloning Kit. Store the LR Clonase™ enzyme mix at -80°C. 
• The pCR™ 8/GW/TOPO™ TA Cloning Kit includes the ready-to-use pCR™ 8/GW/TOPO™ vector, dNTPs, salt solution, water, forward and reverse sequencing primers, control template and PCR primers, One Shot™ TOP10 Chemically Competent E. coli, S.O.C. medium, and a pUC19 control plasmid. Store the Competent Cells at -80°C. Store all other components at -20°C. All components are guaranteed stable for 6 months when properly stored.