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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, andRead more
| Catalog Number | Quantity |
|---|---|
| V47020 also known as V470-20 | 20 Reactions |
Catalog number V47020
also known as V470-20
Price (USD)
1,272.00
20 reactions
Quantity:
20 Reactions
Price (USD)
1,272.00
20 reactions
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-PCR
How 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 a lacZ/gene of interest fusion. This vector features:
•attR sites immediately following the lacZ ORF for fast and efficient recombination with any attL-flanked Gateway™ entry vector to generate the lacZ/target gene fusions
•The CMV promoter for strong expression of the lacZ/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 the lacZ gene so that β-galactosidase will be expressed even if the target fragment being tested contains stop codons
In 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 generating lacZ fusions.
•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
How 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 a lacZ/gene of interest fusion. This vector features:
•attR sites immediately following the lacZ ORF for fast and efficient recombination with any attL-flanked Gateway™ entry vector to generate the lacZ/target gene fusions
•The CMV promoter for strong expression of the lacZ/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 the lacZ gene so that β-galactosidase will be expressed even if the target fragment being tested contains stop codons
In 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 generating lacZ fusions.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
For Use With (Application)RNAi Target Screening
FormatKit
Product Typeβ-Galactosidase Reporter Gene Assay System
Quantity20 Reactions
Reporter GeneBeta-Gal (lacZ)
Sample TypeSynthetic siRNA, RNAi Plasmids (shRNA, miR)
Product LineBLOCK-iT™, Gateway™
PromoterCMV
Unit Size20 reactions
Contents & Storage
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.