just wondering, which quantitation method do you think is the best to use in relative qPCR? Standard curve method (I wonder if someone prefers this because itīs so laborious with standard dilutions on every plate), Pfaffl method (with efficiency determined on several different occasions) or the deltadelta CT method (which I hear is been criticized to be more of an approximation)? Iīm just choosing the method to use; I was advised to use the delta method, but after reading several papers Iīm starting to think Pfaffl would be better. Please let me know your opinions and experiences!
Also: when determining the efficiency, can I just use a dilution of f.e. some of my actual samples? or as I use brain tissue samples, can I determine the efficiency of my primers in a sample from another part of the brain (from which I donīt determine anything and is then "waste")? *get the idea of determining the efficiency but apperently not quite getting the realization *
Thanks alot in advance!!
Edited by rajah, 07 April 2009 - 05:26 AM.