housekeeping gene in real-time PCR - (Jul/24/2006 )

i will work real-time RT-PCR but i get confused some subjects. If you answer me, i am glad. Which stage do you use housekeeping gen?

Beginning, PCR run or after ended Which?

You use your housekeeping gene at all stages. Just treat it like your gene of interest. You set up PCRs with the gene of interest and the housekeeping gene separately (unless mutiplexing) and run them at the same time on the same plate if possible.

Don't think of the housekeeping gene as anything special.

Thank you for interesting. i have another question, i will be contact later.

sincerely you