Protocol Online logo
Top : New Forum Archives (2009-): : PCR, RT-PCR and Real-Time PCR

relative gene expression with multiple reference genes - data analysis (Oct/22/2009 )

Hi friends,
I am doing gene expression profiling of several genes in a plant metabolic pathway. For that I am using three reference genes. How to calculate the expression ratios. I am using light cycler 480 software. Unitll now, I am calculating the expression ratio of a target gene with reference to each reference gene. Is this the correct way or we have to combine the data of all the reference genes, and calculate the expression ratios of target genes. Please explain if you have experience. Thanks

-Linga R Gutha-

We use several reference genes for our work in the lab and we use a program called geNorm to get a normalisation factor. From my understanding the program calculates the average of each sample across the different reference genes and spits out a normalisation number, and using this number your normalise your target gene expression against that to get your fold changes. Hope that helps.