reference gene - (May/12/2011 )
i'm carrying out gene expression experiment on bacteria. i used 16sRNA as a refrence gene, but the Ct value is around 6 while one of my target gene Ct is 27, should i start with a lower cDNA conc. and shift the target gene Ct >30, decrease the primer concentration for the refrence gene or accept the result as it is. does anyone have a clue.
-lilies-
Decrease primer concentration for reference gene. It's called primer limiting. The Ct of your gene of interest and reference gene should be similar. You may try less abundant reference genes.
-Trof-
Trof on Thu May 12 13:09:55 2011 said:
Decrease primer concentration for reference gene. It's called primer limiting. The Ct of your gene of interest and reference gene should be similar. You may try less abundant reference genes.
Thank you, i will try the primer limiting advice
-lilies-