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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Routing Protocol vs Ro 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.
The amounts given are for one well of a 6-well plate. In this protocol, cell plating and transfection are performed on the same day.
Cell strategies for high transfection efficiency | Laboratory Talk
Shortly before transfection, seed 3x cells per well of a 6-well plate in ? Cells may alternatively be seeded after step 3 of this protocol. The amount of transfection reagent and DNA required for optimal performance may vary, depending on the cell line and gene target. Incubate the samples for 10? Add the complexes drop? Gently swirl the plate to ensure uniform distribution of the transfection complexes.
Incubate the cells with the transfection complexes under their normal growth conditions and monitor gene expression after an appropriate time e. Change the medium as required.
The optimal incubation time for gene expression analysis depends tatractene the cell type, the gene expressed, and the method of analysis. This can be determined by performing a time?
For stable transfections, passage cells into the appropriate selection medium attravtene Maintain cells in selective culture medium until colonies appear.
For this reason, further optimization is recommended to achieve maximum transfection efficiency. As a general rule, optimization of transfection can be performed by varying DNA and Attractene Reagent amount as follows: Use the reagent and nucleic acid amounts listed in the protocol.
Cell strategies for high transfection efficiency
The table below shows a pipetting scheme with 5 different conditions we recommend to test when optimizing transfection i. As a starting point, we recommend using 6-well plate? The effect of using greater or lesser amounts of DNA and Attractene Transfection Reagent can be observed using these combinations. 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.
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. Transfection optimization Optimizing DNA 0.