luciferase reporter assay

Scott Brown via methods%40net.bio.net (by SBrown from ccia.unsw.edu.au)
Wed Jul 8 16:18:17 EST 2009


Hi,

Can anyone recommend a protocol for harvesting 293T to be usewd in a luciferase reporter assay. i have no experience with this tecnique, i have one construct with CMV in it and i expect this to be constituatively expressed, i have used this to guage how green the cells are going to be and therefore i have plated serial dilutions of cell # to work out the optimal cell density. I have a promotorless construct as a negative control etc etc
What i am after is any tricks of the trade or advice as to how i go about taking these cells, specifically this step and then any information on how i go about setting up the cells in a white 96well plate to be analysed.
Also can anyone recommend a supplier for the reagents, is promega the standard?

Thanks in advance

Scott

Scott Brown
PhD Candidate
Molecular Carcinogenesis Program
Children's Cancer Institute of Australia for Medical Research
High Street (PO Box 81)
RANDWICK NSW 2031
AUSTRALIA

Phone: +61 2 9382 1829
Fax: +61 2 9382 1850
Email: sbrown from ccia.unsw.edu.au
Web: www.ccia.org.au 
Children's Cancer Institute Australia is the only independent medical research institute in Australia solely devoted to research into the causes, prevention and cure of childhood cancer. Our vision is to save the lives of all children with cancer and eliminate their suffering.
The information contained in this message and any annexure is confidential and intended only for the named recipient(s). If you have received this message in error, please contact the sender immediately and destroy the original message.


Children's Cancer Institute Australia for Medical Research is the only independent medical research institute in Australia devoted to research into the causes, prevention, better treatment and ultimately a cure of childhood cancer. Our vision is to save the lives of all children with cancer and to eliminate their suffering.

The information contained in this message and any annexure is confidential and intended only for the named recipient(s). If you have received this message in error please contact the sender immediately and destroy the original message.



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