TransFect Protocol Database. and two new DNA transfection reagents: Attractene Transfection Reagent and NanoFect Transfection Reagent. I tried Lipofectamine and Attractene which both had a bad efficiency. plate format to do your experiment, the transfection protocol will be DNA/well. 年7月29日 QIAGEN Supplementary Protocol Protocol: Fast?Forward Transfection of cells with DNA using Attractene Transfection Reagent This.
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Forward Transfection of cells with DNA using Attractene Transfection Reagent This protocol uses standard parameters and might require further optimization for transfection with this cell type. Use the reagent and nucleic acid amounts listed in the protocol. To complement and enhance the existing portfolio of trusted transfection solutions, Qiagen has launched the TransFect Protocol Database and two new DNA transfection reagents: Use of the TransFect Database is free of charge and no registration is required.
Directory Resources Events Get Listed. However, for some cell types, especially difficult-to-transfect cells such as suspension cells and primary cells, higher transfection efficiencies may be achieved by plating cells 24 hours prior to transfection for a protocol, consult the Attractene Transfection Reagent Handbook at www. Shortly before transfection, seed 3x cells per well of a 6-well plate in ? Absence of animal-derived components also facilitates regulatory compliance.
As a general rule, optimization of transfection can be performed by varying DNA and Attractene Reagent amount as follows: Home PCR Cell strategies for high tr The instrument range of Hygiena International Ltd, experts in rapid microbial detection, monitoring…. For stable transfections, passage cells into the appropriate selection medium 24?
NanoFect Reagent is completely chemically synthesised, free of animal-derived components, and has been tested for the absence of endotoxins. For this reason, further optimization is recommended to achieve maximum transfection efficiency.
Cell strategies for high transfection efficiency
If you require a thermal cycler that delivers high performance and high throughput, while still ret…. The Fast-Forward Protocol, in which cell plating and transfection are performed on the same day, has been shown to work successfully with a wide variety of cell types.
The effect of using greater or lesser amounts of DNA and Attractene Transfection Reagent can be observed using these combinations. The amounts given are for one well of a 6-well plate. Hygiena expands with PCR-based systems to cover the entire contamination detection spectrum Added: Make money from your old lab equipment. This reagent is suitable for a range of transfection applications, including transient or stable transfection, transfection of shRNA short-hairpin RNA vectors, and DNA cotransfection.
Incubate the samples for 10? In this protocol, cell plating and transfection are performed on the same day. Transfection optimization Optimizing DNA 0. Rather than adapting existing protocols to fit the cell type, the TransFect Database enables researchers to access exactly the protocol needed by simply entering the cell type, nucleic acid, and plate format into the online system. Incubate the cells with the transfection complexes under their normal growth conditions and monitor gene expression after an appropriate time e.
Cell strategies for high transfection efficiency | Laboratory Talk
In addition, the absence of lipids makes this reagent suitable for lipid or signal transduction research. As a starting point, we recommend using 6-well plate? Add the complexes drop? The latest reagents use advanced technology to meet all DNA transfection needs.
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Cells may alternatively be seeded after step 3 of this protocol. Cell strategies for high transfection efficiency 19 Feb by Qiagen.
Gently swirl the plate to ensure uniform distribution of the transfection complexes. Simple Safe Parallel Reaction Sampling.
Maintain cells in selective culture medium until colonies prrotocol. For these reasons, it is ideal for use when absence of animal-derived components is a priority, for example, in biopharmaceutical applications. Using Attractene Reagent also ensures exceptionally low cytotoxicity.
The TransFect Protocol Database is an invaluable resource which provides cell-specific transfection protocols. The amount of transfection reagent and DNA required for optimal performance may vary, depending on the cell line and gene target. Ease of handling means that Attractene Aftractene is suitable for use with automated systems.
Prime thermal cyclers provide flexible PCR Added: Change the medium as required. This can be determined by performing attrcatene time? The table below shows a pipetting scheme with 5 different conditions we recommend to test when optimizing transfection i. Routing Protocol vs Ro The optimal incubation time for gene expression analysis depends on the cell type, the gene expressed, and the method of analysis.