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- A single construct for $85
- 4 constructs for only $275!
- Guaranteed knockdown for any gene
- Every construct with a GFP reporter
- Less than the material cost, why do it yourself?
Simply provide us with your gene's accession number for any target gene or siRNA target sequence and we will create the lentivirus siRNA construct for you within 2 weeks!
Once we get your gene name, we will search our siRNA database of over 20,000 publications covering over 7,000 human and mouse genes. Then we will design four constructs with high knockdown efficiency against the target. If your gene of interest is not yet published, we will design four target sequences using our internal siRNA software. With four siRNA constructs, at least one of the constructs should give over 70% gene knockdown efficiency in appropriate target cells with over 70% transfection efficiency.
We also have a scramble construct in our proprietary vector as the ideal control for your experiments.
- siRNA in lentiviral vectors for either transfection or viral infection.
- iLenti™ vectors are very stable, no special competent cells are required.
- iLenti™ uses more potent 27-29 bp oligos.
- iLenti™ uses convergent promoters to avoid hairpin loop structures, making sequencing and plasmid growth much easier.
Refer to the paper "Transgenes Delivered by Lentiviral Vector are Suppressed in Human Embryonic Stem Cells in a Promoter-Dependent Manner" by Xia et al. in Stem Cells and Development 16:167-176 (2007).
EGFP expression is dependent on promoter strength, which may differ between different cell types. The promoter for the EGFP gene is from CMV. Even though the promoter is ubiquitous, the expression of EGFP may be suppressed in some target cells. That does not necessarily mean that the vector is not in the cells. The vector may be there and the siRNA may be expressed because the promoters for the siRNA are different.
You can try Western blotting against the target gene product along with a standard curve to evaluate whether siRNA is being expressed and functional in the target cells.
Lentiviral vector stocks are tested for the presence of replication-competent lentivirus (RCL) by monitoring p24 antigen expression in the culture medium of transduced 293T cells for 30 days. Serial passaging of the transduced p24 cells over this period allows for the amplification of RCL. Since the vectors are replication-defective, no amplification of p24 signal would normally be expected. However, if there were an increase in P24 signal over time, it would be indicative of RCL contamination. The p24 antigen is assayed with a commercial kit from http://www.zeptometrix.com/ . This assay has been performed on multiple vector lots without a positive result.
U6 forward sequencing primer
You can use the H1 Primer to sequence in the reverse direction if required:
H1 sequencing primer: 5'-CAACCCGCTCCAAGGAATCG-3'