siRNA/miRNA high throughput screen service
Large-scale siRNA screens are used to uncover genes that are involved in particular cellular pathways, and have become an indispensable research tool in all molecular life sciences. GenePharma offers state-of-the-art high-throughput siRNA screening to identify the functions of thousands of genes, and promptly identify new research targets for our customers.
Available siRNA libraries for high-throughput screening:
- Full human genome siRNA library (21585 genes).
- Full human miRNA library and inhibitor (983×2).
- Cell cycle siRNA library (131 genes).
- Deubiquitinating enzymes siRNA library (127 genes).
- Phosphorylase library (257 genes).
- Cytokine receptor siRNA library (319 genes).
- G-protein-coupled receptor siRNA library (516 genes).
- Apoptosis siRNA library (558 genes).
- Protein kinase siRNA library (779 genes).
- Custom siRNA library (100-2000 genes).
The service procedure for our high-throughput siRNA screens is as follows:
1. Please fill in and submit the on-screen application form. You also need to provide background information about which cell type and which cellular pathways are of interest.
2. GenePharma's scientific staff will design a plan for the siRNA screen based on your requirements and information.
3. Our researchers will optimize the screen conditions based on the information that the clients provide (i.e. cell type, migration character, etc.). We will carefully specify the transfection conditions to the target cell line to optimize the siRNA transfection efficiency, as this is a key factor for a successful siRNA screen. To do so, we perform a quantitative evaluation.
Firstly, we experimentally select dependable siRNA controls: a strong positive control and a stable negative control that behave predictably between plates and different experiments.
Secondly, we conduct a high-throughput screening with positive and negative controls on the same plate every time. Thirdly, to select the positives, we first analyze the distribution of values according to normal distribution, then we normalize the values and find out those that greatly deviate from the list. We perform the primary screening by a pool of 3 siRNAs. For these positives, the validation will be done by testing the individual siRNA of each gene. It is better to verify the result with a second cell assay.
4. Once we have optimized the transfection conditions using the proper controls, we perform a small-scale trial screen: we will use 354 well plates or self-assembled cell microarray (SAMcell) chips to screen about 100 genes and testify whether the result is reliable.
5. We conduct the large-scale siRNA library screen on the target genes. The screening will be performed using Multi-well plates (384 well or 96 well), or on our patented self-assembled cell microarray (SAMcell), in which 100 siRNAs are microprinted on a single cover-chip . [1.] Furthermore, we developed a unique approach for manufacturing esiRNAs (endoribonuclease-prepared siRNA). [2.]
6. After the experimental part of the large-scale library screen, we will statistically analyze the data and provide a detailed report to the client. Together we discuss and decide on what next steps are possible.
The whole screening process, from design till final report and discussion, lasts about 3-6 months, depending on the scale of the library and the screening plans. Due to the scope of the project, an advance payment is customary.
For more information about screen service costs, please contact bd@genepharma.com. For a detailed description of our procedure and equipment, please check the siRNA screen guide manuals, or contact our scientific staff (support@genepharma.com). For high impact references click here.
[1.] Zhang H. et al., Nature Communication 2011 v2:554; Rong Y. et al., Nature Cell Biology 2012 v14(9):924-34.
[2.] Huang H. et al. Lab Chip, 2011; Wang Z. et al. PLoS ONE, 2012. |
