Bio-Rad Firmware & Software Updates - Manual (Page 118)

MiniOpticon Instruction Manual
107
Standard Deviation of Relative Quantity
The standard deviation of the relative quantity is calculated with the following formula:
Where:
• SD Relative Quantity = standard deviation of the relative quantity
• SD C
q
sample = Standard deviation of the C
q
for the sample (GOI)
• Relative Quantity = Relative quantity of the sample
• E = Efficiency of primer and probe set. This efficiency is calculated with the formula
(% Efficiency * 0.01) + 1, where 100% efficiency = 2
• GOI = Gene of interest (one target)
Normalization Factor
The denominator of the normalized expression equation is referred to as the normalization
factor. The normalization factor is the geometric mean of the relative quantities of all the
reference targets (genes) for a given sample, as described in this formula:
Where:
• RQ = Relative quantity
• n = Number of reference targets
• GOI = Gene of interest (one target)
Efficiency Corrected Cq (Cq
E
)
The efficiency corrected Cq is calculated by the following formula:
Where:
• E = efficiency
Mean Efficiency Corrected Cq (MCq
E
)
The mean efficiency corrected Cq is calculated with the following formula:
Where:
• Cq
E
= Efficiency corrected Cq
• n = number fo replicates
SD Relative Quantity SD Cq
GOI
Relative Quantity
Sample X
Ln (E
GOI
  
=
Normalization Factor
sample (GOI)
(RQ
sample (Ref 1)
RQ
sample (Ref 2)
 RQ
sample (Ref n)
)
  
1
n
--
=
Cq
E
Cq (log(E)/log(2))

=
MCq
E
Cq
E (Rep 1)
Cq
E (Rep 2)
 Cq
E (Rep n)
+ + +
n
---------------------------------------------------------------------------------------------------------- =