( aspirate the growth medium fr...)Transfection Reagent Selector Kit Handbook

aspirate the growth medium from all wells and wash cells once with 2 ml PBS. To wells 111 add 1600 l fresh growth medium (can contain serum and antibiotics). To wells 1222 no medium should be added. For suspension cells, skip this step.

Add 600 l of growth medium (can contain serum and antibiotics) to all tubes. Mix by pipetting up and down twice, and immediately add the total contents drop-wise onto the cells in the appropriate wells. Add the contents from tube 1 to well 1, from tube 2 to well 2, from tube 3 to well 3, and so on. Gently swirl each dish to ensure uniform distribution of the complexes.

Incubate both Effectene Reagent plates (wells 111) at 37C and 5% CO₂ to allow for gene expression.
Incubation length is determined by the assay and gene used. Experiments have shown that in many cases removal of transfection complexes is not necessary.

Incubate both SuperFect Reagent plates (wells 1222) for 23 h at 37C and 5% CO₂ to allow for uptake of the transfection complexes. After this time, remove the medium containing the remaining complexes from the cells by gentle aspiration, and wash cells 1x with 2 ml of PBS. Add fresh growth medium (can contain serum and antibiotics). For suspension cells, removal of transfection complexes is usually not necessary.
If cytotoxicity is observed with suspension cells, remove the transfection complexes by centrifugation after a 23 hour incubation period. Remove the medium from the cells, resuspend cells in fresh medium containing s
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
Related biology technology :

1. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell Line
2. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
3. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
4. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
5. Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance
6. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
7. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
8. High-Efficiency Transfections Achieved with New Low-Toxicity Reagent
9. Efficient Transfection of Neurospora Crassa
10. Eppendorf Multiporator Transfection Protocols for Eukaryotic Cells
11. Efficient Transfection of Neurospora Crassa