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shRNA clone sets

Answer: All you need to do is go to the shRNA clones search page, search for your gene, and then choose the appropriate clones that will work for your system. If you have any custom requirements, then you will need to contact us and, after determining what you need, we will send you a custom quote.
Answer: GeneCopoeia provides a set containing 3 individual shRNA constructs and a scrambled control. Each clone is provided as of 5 µg purified plasmid. A datasheet containing information for each clone, vector and restriction enzyme sites is available for download on our website. You will need a GeneCopoeia account, your sales order number, and your catalog number(s) in order to download the datasheet.
Answer: GeneCopoeia guarantees that the shRNA sequences in the expression cassettes are identical to the target gene. If none of the four constructs produce a 70% or greater knockdown efficiency as determined by qRT-PCR, and the inefficiency is caused by a flaw in our construct design, then we will provide another set of four new clones targeting the specific gene free of charge.

Answer: We offer both promoters so customers can choose based on their preference. U6 is the stronger promoter, however, because it is stronger there is a greater chance of causing off-targeting effects or cellular toxicity. In most cases either promoter should be fine to use, however, there are tissue and cell specificities associated with each promoter. We recommend doing a literature search to see what other researchers have used for the particular cells you are working with.


Answer: Yes, Stbl3 is recommended.
Answer: You should measure the knockdown efficiency when the transfection efficiency is greater than 80%. The reporter gene in the vector is used to monitor transfection efficiency. Alternatively, if the transfection efficiency of your cell line is low, you can order the lentiviral versions of your clones and transduce your cells with the lentiviruses. RNA can be harvested from transfected cells and used in quantitative RT-PCR to estimate the reduction in gene expression. Western blot is recommended over qPCR to evaluate the silencing effect of the shRNA construct(s). Gene expression levels from cells transfected with a scrambled control clone must be compared with the shRNA transfected samples.
Answer: Factors influencing transfection efficiency include 1) The quality of the plasmid DNA; 2) The condition of the cells; 3) The quality, condition, or age of your transfection reagents and plasmids; 4) The cell density at the time of transfection; and 5) The contact time between cells and the DNA/transfection reagent complex.
Answer: The scrambled control clone is constructed by cloning a scrambled sequence (one that does not match any genomic sequences) into shRNA vectors. It serves as a negative control to eliminate the potential non-specific effect induced by expression of the plasmid.
Answer: We strongly recommend performing a kill curve on each batch of cells to determine the optimal puromycin concentration.