Product and Protocol Overview:

1. Four pre-designed shRNA plasmids are provided with each purchase. One negative control shRNA is also provided.

2. Transform provided plasmids separately into competent E. coli cells.

3. Pick colonies for large-scale high-quality transfection-grade stock plasmid preparation.

4. Transfect plasmids separately into replicate wells of your cell line of interest.

5. Track and characterize transfected cells by fluorescence microscopy, OR enrich the transfected cell population by FACS or by selection.

6. Assay the extent of knock-down in enriched or selected cells by real-time PCR.

7. Perform your biochemical, cell or molecular biological gene function assay.

Applications:

1. Use cell lines with either low or high transfection efficiencies. Study gene function in virtually any cell line.

2. Use the neomycin resistance marker to select for stably transfected cells. Study slower responses to RNA Interference using any biochemical, cell or molecular biology gene function assay.

3. Use the GFP marker to identify and track transiently transfected cells by fluorescence microscopy. Study quick responses to RNA Interference using cell-based imaging gene function assays such as immunofluorescence or morphology.

4. Use the GFP marker to enrich for transiently transfected cells by Fluorescence Activated Cell Sorting (FACS). Study quick responses to RNA Interference using any biochemical, cell or molecular biology gene function assay.