Protocol Online logo
Top : Forum Archives: : Transfection and Transduction

Harvesting cells following transfection - Scrapping or trypsin (Nov/14/2007 )

I have been conducting a series of transient transfections in mouse aorta endothelial cells by microporation and assaying transfection efficiency using Luciferase and ß-gala reporters. I have been harvesting the cells by scrapping but was wondering whether I could simply use trypsin followed by washing to harvest the cells.

I know it is a very basic questions, but I have been told the trypin will interfer with assaying luciferase and ß-gala, and would like to use this approach rather than scrapping as to reduce time and increase the number of samples I can process; I have alarge time course study to perform and would ultimately like to get everything done within 24hrs without staggering everything too much.

All comments greatfully received.



Why not use lysis buffer that is compatible with the assays?


We bought a luciferase kit for assay and they had supplied us with the buffer for extraction.