AllStars Hs Cell Death Control siRNAFor siRNA transfection optimization and positive control experiments in human cells
AllStars Hs Cell Death Control siRNA is a blend of highly potent siRNAs targeting ubiquitously expressed human genes that are essential for cell survival. Knockdown of these genes induces a high degree of cell death, which is visible by light microscopy. Transfection efficiency can be quickly estimated by simply observing cells by straightforward light microscopy 48–96 hours after transfection of AllStars Hs Cell Death Control siRNA, avoiding the need for any complex or laborious downstream assay. AllStars Hs Cell Death Control siRNA is a useful tool for siRNA transfection optimization and can be used routinely as a positive control.
Performance
Fast and easy detection in a wide range of cell typesThe performance of AllStars Hs Cell Death Control siRNA is validated for a wide range of cell types (see table).
Highly efficient knockdown and cell deathKnockdown efficiency using AllStars Hs Cell Death Control siRNA has been quantified by real-time RT-PCR and is >95% for each target mRNA. The AllStars Hs Cell Death Control significantly outperforms other commercially available toxic control siRNAs in both the level of cell death induced and in the wide range of cell types in which it has been validated. Consistently high levels of cell death are achieved using AllStars Hs Cell Death Control, ensuring effective control experiments (see figure High level of cell death).
Principle
Easy transfection optimizationWhen establishing RNAi in start-up experiments or in a new cell line, multiple transfections should be performed under different conditions to determine the optimal conditions for maximum transfection efficiency. These experiments can be performed using AllStars Hs Cell Death Control siRNA (see figures Easy siRNA transfection optimization and Cell death in primary cells). Transfection conditions that result in the greatest degree of cell death compared with transfection with a nonsilencing control siRNA can be maintained in future experiments. Reliable high-throughput RNAiIn high-throughput RNAi screens, the use of internal positive controls is essential to ensure that optimal transfection conditions are maintained on every plate of the screen. Positive controls that can be analyzed phenotypically have the advantage of allowing transfection efficiency to be observed immediately. Using AllStars Hs Cell Death Control siRNA on every plate allows rapid and inexpensive estimation of the transfection rate on individual plates.
Procedure
Using AllStars Hs Cell Death Control siRNA, transfection efficiency can be quickly estimated by simply observing cells by straightforward light microscopy 48–96 hours after transfection, avoiding the need for any complex or laborious downstream assay. AllStars Hs Cell Death Control siRNA can be ordered in tube format in 1 nmol, 5 nmol, or 20 nmol amounts. This control is also available in 96-well plates and 384-well plates (FlexiPlate siRNA) in 1 nmol, 0.25 nmol, or 0.1 nmol amounts.
Applications
|