Bio-Rad Firmware & Software Updates - Manual (Page 145)

Gene Expression Analysis
132
Corrected Values Formulas
A difference between corrected values and noncorrected values is seen only if a standard
curve is created as part of the real-time PCR run. The software uses three equations in
determining the error propagation:
• Standard Error
• Standard Error for Normalized Expression
• Standard Error for the Normalized Gene of Interest (target)
The formula for standard error is shown here:
Where
• n = Number of reference targets (genes)
• SD = Standard deviation
The standard error for the normalization factor in the normalized expression formula is shown
here:
Where:
• n = Number of reference targets
• SE = Standard error
• NF = Normalized expression
• RQ = Relative quantity
The standard error for normalized gene of interest (GOI) formula is shown here:
Where:
• SE = Standard error
• GOI = Gene of interest (one target)
• NF = Normalization factor
• n = Number of reference targets
Standard Error
SD
n
-------- =
SE NF
n
NF
n
SE RQ
sample (Ref 1)
n SE RQ
sample (Ref 1)
×
---------------------------------------------------------
 
  2 SE RQ
sample (Ref 2)
n SE RQ
sample (Ref 2)
×
---------------------------------------------------------
 
  2
…
SE RQ
sample (Ref n)
n SE RQ
sample (Ref n)
Ч
---------------------------------------------------------
 
 
2
+ + + Ч =
SE GOI
n
GOI
n
SE NF
n
NF
n
----------------  
  2
SE GOI
GOI
-----------------  
 
+
2
Ч =