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RNAi: The Controls You Need


Ambion scientists use and recommend a number of different controls for siRNA experiments. Most of these coincide with the suggested controls detailed in an editorial published in Nature Cell Biology [1]. Here we outline these various controls and discuss the importance of each in RNAi experiments.

Recommended Controls and How to Accomplish - Click for larger version


Scrambled (Negative) siRNA Control

Comparing cells transfected with a nonsense or scrambled sequence siRNA control to nontransfected cells can reveal changes caused by siRNA delivery. Silencer Negative siRNA Control #1 is a validated negative control that has limited sequence similarity to the human, mouse, or rat genomes. Ambion scientists, as well as many other scientists around the world, routinely use this negative control in their RNAi experiments.


Positive siRNA Control

A positive siRNA control should be used in experiments to monitor and optimize siRNA transfection efficiency. When the positive control fails to elicit the expected reduction in gene expression, poor transfection is immediately suspected. Ambion's Silencer GAPDH siRNA is a commonly used positive control because GAPDH is expressed at easily detectable levels in most cell types.


Multiple siRNAs to a Single Target

There are several example
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Related biology technology :

1. RNAi: A How To for New Users
2. RNAi: Get the Whole Story
3. RNAi: Size Does Matter
4. Housekeeping Genes: Universal Positive Controls in siRNA Knockdown Experiments
5. Designing Controls for siRNA Experiments
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