Applied Biosystems 7900HT User Manual page 356

Appendix D Theory of Operation
D-10 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
September 1, 2004 11:39 am, App_Theory.fm
Rearranging gives the following expression:
∆
–
C
× ( ) T
X = K 1 + E
N
The final step is to divide the X
X
× 1 ( E )
N q , K +
=
-------------- - --------------------------------------------- -
X
,
× 1 ( E )
N cb
K +
where:
∆∆C = ∆C – ∆C
T T,q
For amplicons designed and optimized according to Applied Biosystems guidelines
(amplicon size <150 bp), the efficiency is close to 1. Therefore, the amount of target,
normalized to an endogenous reference and relative to a calibrator, is given by:
∆ ∆
– C
T
2
DRAFT
for any sample q by the X
N
∆
– C
T q ,
∆ ∆
– C
1 ( E ) T
=
+
∆
–
C
,
T cb
T,cb
for the calibrator (cb):
N