RNAi Workflow Overview

The GeneAssist Workflow Builder will walk you through the RNAi workflow, making it simple to find the reagents you need to make each step successful.

Below is an overview of the RNAi workflow. The tabs in the GeneAssist Workflow builder correspond to each step in the below workflow. At each tab, you can click the

button at the top right to get information about that particular step in the workflow. The

icon leads to information about how to use the tool to select the reagents you need for your research.

Step 1. Obtain effective siRNAs/gene expression assays

To obtain gene silencing, potent and specific siRNAs must be designed. Effective siRNA design and rigorous quality control of synthesis and purification will prevent the waste of both time and money.

TaqMan® Controls

TaqMan®

siRNA Controls

siRNA

Step 2. Optimize siRNA delivery

Efficient, reproducible siRNA delivery is essential for successful RNAi experiments. Because siRNA delivery is so critical, it is highly worthwhile to invest the time and effort to determine the best siRNA delivery method and conditions for each cell line studied.

Optimization

Electroporation

Transfection

Step 3. Test siRNA silencing efficiency

Once siRNAs are obtained and delivery is optimized, the next step in a typical gene silencing experiment is to assess the silencing efficiency of the selected siRNAs.

qtPCR

RNA Isolation

Step 4. Examine biological impact of silencing

The ultimate goal of most RNAi experiments is to understand the biological impact of the reduced target gene expression.

Immunodetection

qtPCR

Cell Based Assays

Step 5. Correlate silencing efficiency to biological effect

In order to fully understand the biological impact of reduced target gene expression, it is necessary to correlate the observed phenotype with the level of knockdown induced.

Immunodetection

qtPCR

RNA Isolation