BIO RAD CFX384, Instruction Manual

Page: 110 / 158

CFX96 and CFX384 Systems Manual
101
9
Gene Expression Analysis
Read this chapter for information about performing Gene Expression Analysis:
• Gene Expression (below)
• Plate setup for gene expression analysis (page 102)
• Gene Expression tab (page 102)
• Experiment Settings window (page 107)
• Gene Study (page 107)
• Gene Study Report window (page 113)
• Gene expression calculations (page 115)
Gene Expression
With the use of stringently qualified controls in your reactions, you can run a gene expression
experiment to normalize the relative differences in a target concentration between samples.
Typically, message levels for one or more reference genes are used to normalize the
expression levels of a gene of interest. Reference genes take into account loading differences
or other variations represented in each sample, and they should not be regulated in the
biological system being studied.
Open the Gene Expression tab to evaluate relative differences between PCR reactions in two
or more wells. For example, you can evaluate relative numbers of viral genomes, or relative
number of transfected sequences in a PCR reaction. The most common application for gene
expression study is the comparison of cDNA concentration in more than one reaction to
estimate the levels of steady state messenger RNA.
The software calculates the relative expression level of a target with one of these scenarios:
• Relative expression level of a target sequence (Target 1) relative to another target
(Target 2). For example, the amount of one gene relative to another gene under the
same sample treatment
• Relative expression level of one target sequence in one sample compared to the
same target under different sample treatments. For example, the relative amount of
one gene relative to itself under different temporal, geographical, or developmental
conditions