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RNAi Screening service
Using siRNA screening to select the genes which related cells life cycle has become a common tool for the research of gene chemistry and life science. The high throughput screening could analyze the functions of thousands genes, and promptly identify the relative genes,thus find new research targets
 

siRNA/miRNA high throughput screen service 
Using siRNA screening to select the genes which related cells life cycle has become a common tool for the research of gene chemistry and life science. The high throughput screening could analyze the functions of thousands genes, and promptly identify the relative genes,thus find new research targets. 

The siRNA libraries available: 


1)full human genome siRNAlibrary (21585 genes) 
2)full human miRNA library and inhibitor (983×2) 
3)cell cycle siRNAlibrary (131 genes) 
4) Deubiquitinating enzymes siRNA library (127 genes) 
5) phosphorylase library (257 genes) 
6) cytokine receptor siRNA library (319 genes) 
7) G-protein-coupled receptor siRNA library (516 genes) 
8) apoptosis siRNA library (558 genes) 
9) protein kinase siRNA library (779 genes) 
10)custome siRNA library (100-2000 genes)

service procedures:

1.Raise the opinions of siRNA screen: clients first land our web to understand our service content, and fill in the screen application form, and send it back to us, the clients also need to provide the background information of your target cell pathways. 
2.confirm the screen plan. GenePharma's research staffs will establish the screen plan based on the clients information. Clients will sign the contract with GenePharma and pay some proportion of front money. 
3.optimize the screen condition. We will optimize the screen condition based on the information that the clients provided( cell type, migration character,so on) 
cell transfection efficiency optimize is the key factor to conduct a success siRNA screen。 
3.screen modle quantitive evaluation and optimize
First of all, find out a strong positive control and a stable negative control. The values of positive control and negative control should be stable between plates and different experiments. So that the value of Z' could be analyzed and optimized to judge whether those two controls is suitable enough to conduct following large scale screening






Second, conduct high-throughput screening with positive control and negative control on the same plate every time.
Third, to select the positives, we first analyze the distribution of values accord to normal distribution, then we normalize the values and find out those greatly deviates 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.small scale trail screen, once confirmed the cell screen condition, we will use the 354 well plates or SAMcell chips to screen about 100 genes and testify whether the result is reliable. 
5、.large scale siRNA library screen: TO conduct massive siRNA/miRNA screen on the target genes. 
6、apply the screen report; after the massive screen, we will statistic and analyze the data, provide report to the clients, and descide the next steps。 
7、 for the positive results, we will contact with the clients, and testify the positive results by chery pick-up method. 
The whole screen process last about 3-6 months, the time will descide the scale of the library and screen plan.
Screen Technique :

1. The screening will be performed based on Multi-well plates (384 well or 96 well), or on self-assembled cell microarray (SAMcell) (patented), in which 100 siRNAs are microprinted on a single covership. (Zhang H. et al. Nature Communication 2011 v2:554; Rong Y. et al, Nature Cell Biology 2012 v14(9):924-34). 
2. We developed a unique approach for manufacturing esiRNA (Huang, H et al. Lab Chip, 2011; Wang, Z. et al. PLoS ONE, 2012).

For more information please check the siRNA screen guid manuals.
Our success siRNA screen examples










EnSpire® Multimode Plate Readers (PerkinElmer).
Success examples
We finished the screening of functional genes regulating cell migration. The library we screened includes miRNA mimics libraries (~900) (Zhang H. et al. Nature Communication 2011 v2:554.) and Human kinase siRNA library (~750) (under review).
We finished the siRNA screening of functional genes regulating autophagosome formation. The library we screened includes custom-based siRNA library (~300) (Rong Y. et al, Nature Cell Biology 2012 v14(9):924-34). The screen using the whole human genome siRNA library (ABI, ~20,000) is undergoing.
We finished the screening of functional genes regulating cell invasion by use of multi-well plates. The library we screened includes miRNA mimics libraries (~900) and human kinase siRNA library (~750).
We finished the screening of functional genes regulating cell apoptosis. The library we screened includes miRNA mimics libraries (~900), Human kinase siRNA library (~750) and esiRNAs(~100) (Wang Z. et al. PLoS ONE, 2012)
We accomplished the screening of functional genes regulating mitochondrial fission and fusion. The library we screened includes miRNA mimics libraries (~900) (paper in preparation).
We are screening functional genes regulating NADH metabolism. The library includes Human kinase siRNA dibry (~750).
We are screening functional genes capable of generating synthetic lethal with a chemical. The library includes Human kinase siRNA library (~750) and phosphatase siRNA library (~300).
We are performing the screen in other cell assays, including insulin resistance, modulators in TGF-beta pathway, etc.

 
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