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SHXC01 Sigma

MISSION® LentiExpress Optimization Plate

Optimization of cell number and/or MOI for LentiExpress Kinases

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Properties

Related Categories Functional Genomics and RNAi, LentiExpress Plates, MISSION shRNA, Molecular Biology
storage temp.   −70°C

Description

General description

Lentiviral systems are commonly used for the efficient delivery of shRNAs to a broad variety of cell types. Optimization of transduction conditions is a central component of successful delivery using this approach. For maximal transduction and knockdown, it is important to determine the optimal ratio of virus to cells, or the MOI (multiplicity of infection), for each new cell type. The MISSION LentiExpress Optimization Plate accelerates this process allowing rapid determination of the optimal cell density for transduction. The 96-well plate contains pre-arrayed aliquots of TurboGFP Control Transduction Particles, allowing the user to simply add the desired numbers of their cell type of interest. Efficiency of transduction is subsequently monitored by assessment of GFP fluorescence.

The TurboGFP Control Transduction Particles serve as a useful positive control in experiments using the MISSION shRNA library clones. TurboGFP is an improved variant of the green fluorescent protein copGFP cloned from the copepoda Pontellina plumata. The TurboGFP transduction particles are produced from the sequence-verified lentiviral plasmid, pLKO.1-puro-CMV-TurboGFP, Catalog Number SHC003. The TurboGFP sequence is delivered to cells by replication incompetent lentiviral particles. Unlike murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the specific shRNA sequence into differentiated and non-dividing cells, such as neurons and dendritic cells. Self-inactivating replication incompetent viral particles are produced in packaging cells by co-transfection with compatible packaging plasmids.

In addition, the lentiviral transduction particles are pseudotyped with an envelope G glycoprotein from Vesicular Stomatitis Virus (VSV-G), allowing transduction of a wide variety of mammalian cells.

Linkage

To view the data and protocols for the LentiExpress kinase panel visit the links below.
Protocols
Applications

Other Notes

Adding a range of cells for the transduction step allows a quick determination of the optimal cell density and transduction efficiency for cell types of interest. The product is intended for optimization of cell number and/or MOI for the LentiExpress Kinases product, Catalog No. SHX001.

Lentiviral Particles for the LentiExpress Optimization Plate are pre-diluted to approximately 5,000 viral particles per well in a single reaction volume of 30μL in 96-well cell culture plates.

Packaging

MISSION LentiExpress Optimization Plate consists of 80 wells containing 30 μL of TurboGFP control or control shRNA Lentiviral Particles. Sixteen wells contain 30 μL of media only.

Legal Information

Use of this product is subject to one or more license agreements. For details, please see http://sigmaaldrich.com/missionlicense .

LentiExpress is a trademark of Sigma-Aldrich Co. LLC

MISSION is a registered trademark of Sigma-Aldrich Co. LLC

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Safety & Documentation

Safety Information

RIDADR 
UN3373 - class 6.2 Biological substance, Category B
WGK Germany 
3

Documents

Certificate of Analysis

Protocols & Articles

Protocols

Optimal Cell Density Determination for Transduction Using MISSION LentiExpress Optimization Plate

Day 0 0.1 Thaw LentiExpress optimization plate at room temperature while completing steps 0.2 through 0.7. 0.2 Fill and label four 1.5 ml tubes with 300 µl of complete media. 0.3 Trypsinize KHOS cell...
Keywords: Microscopy, PAGE

Peer-Reviewed Papers
15

References

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