The Choice of Housekeeping Gene in Quantitative PCR Data Analysis

• Activity Number: 254
• Type: Contributed
• Date/Time: Monday, August 1, 2011 : 2:00 PM to 3:50 PM
• Sponsor: Section on Quality and Productivity
• Author(s): Yi Guo*+ and Michael Pennell and Soledad Fernandez and Dennis Pearl and Thomas Knobloch and Christopher Weghorst
• Companies: University of Florida and The Ohio State University and The Ohio State University and The Ohio State University and The Ohio State University and The Ohio State University
• Keywords: quantitative PCR ; Housekeeping genes ; paired t-tests

Abstract

Real time polymerase chain reaction (quantitative-PCR or Q-PCR), a method of quantitatively measuring gene expression, is widely used in biomedical research. It is a highly sensitive technique which can accurately measure the presence of even trace amounts of mRNA in the cell. To produce reliable results, it is essential to use stably expressed housekeeping genes for data normalization. In this study, a q-PCR dataset that contains 12 cancer related genes from 16 different tumor tissues and 3 housekeeping genes was analyzed. Black raspberry was used as a treatment against cancer and its effects on the expression of these 12 target genes were examined using a paired design. The objective of this study was to examine the choice of housekeeping gene (HKG) on the identification of differentially expressed genes in Q-PCR experiments. Data analysis showed that results differed with choice of HKG since 2 of the 3 HKGs were differentially expressed across treatment groups.