Views
Section7:Transfection Cationic Matrix Reagents
From Assay Guidance Wiki
Superfect (Qiagen), Lipofection, Fugene 6 (Boehringer), Transfectam (Promega), TransFast (Promega), Tfx (Promega), CLONfectin (ClonTech), Lipofectamine Plus (GIBCO)
Once the optimal transient transfection method and cationic matrix reagent have been selected then additional factors can be considered in developing a method for optimizing a transient transfection protocol.
|
Day |
Protocol |
Variables |
|---|---|---|
|
1 |
Seed cells |
Cell number is dependent on cell type |
|
2 |
Transfect cells |
Protocol of transfection is dependent on reagents used (see list above on transfection reagents) |
|
3,4 or 5 |
Determine transfection efficiency, end product or reporter expression |
Depending on the cell type the assay for gene expression is usually measured between 24 to 72 hours after transfection. Induction or stimulation is usually done 48 hours after transfection, and then determine expression levels. Method for determining expression will also depend on type of expression- GFP can be monitored by visual inspection; luciferase is measured by cell lysis, addition of substrate and measurement of light output. |
