qRT-PCR was conducted by using the SYBR Green PCR master mix (Applied Biosystems, CA).
Nested PCR was performed by using the HotStarTaq Master Mix Kit (QIAGEN).
Electrophoresis was implemented at a constant voltage of 170 V in 0.5× Tris-acetate ethylenediaminetetraacetic buffer for 75 min. The gels were visualized by using the Image Master VDS (Pharmacia Biotech, Puurs, Belgium).
PCR conditions included an initial denaturation at 95°C for 15 min for activation of the HotStarTaq DNA polymerase (QIAGEN GmbH, Hilden, Germany), followed by 30 cycles of 94°C for 30 sec, 52°C for 30 sec, and 72°C for 1 min, followed by an extension at 72°C for 7 min. Reactions were performed in 25 μL by using the HotStarTaq Master Mix (QIAGEN GmbH).
Then, the reaction mechanism and pre-exponential factor were analyzed by using the generalized master-plots method.
For a master-slave system, the synchronization means that the state of a slave system follows the state of a master system asymptotically by using the output of the master system to control the slave system.
TaqMan analysis were realized by using the FastStart DNA Master Hybridization Probes kit (Roche Diagnostics), with the following amplification program: 10 min – 95°C for activation of the enzyme, 40×[95°C – 10 sec; 60°C – 10 sec; 72°C – 10 sec].
Q-PCR was performed on a CFX-96 (Biorad) thermocycler by using the 2x SsoFast Master Mix (Bio-Rad Laboratories).
The Megaplex RT product (2.5 μl) was preamplified by using the TaqMan PreAmp Master Mix (Applied Biosystems) and preamplification primers in a 25-μl PCR reaction.
Control is designed by using the relevant variables of master and slave systems.
Analysis with SYBR green technology were realized by using the LightCycler FastStart DNA Master Mix SYBR Green I kit (Roche Diagnostics, Meylan, France).
