Mastercyclers ep realplex4

Total RNA was isolated from the tissues were using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions as previously described (14, 41) . Real-time PCR was performed using the SYBR ExScript PCR kit (Takara Biochemicals, Dalian, China) in a total volume of 10 μL. The primers used in the real-time PCR are shown in Table 1. PCR amplification and quantification were performed using an Eppendorf MasterCyclers ep RealPlex4 (Wesseling-Berzdorf, Germany) as described in our previous report (40) . The data were normalized to the amount of GAPDH.

The reverse transcriptase-polymerase chain reaction (RT-PCR) was performed by using SYBR ® Green Realtime PCR Master Mix (TOYOBO) in a total volume of 10 μl. The primer sequences for Bmal1, Per1, Per2, Cry1, and Rev-erbα were the same as in our previous report [12, 30] . The primer sequences for Cck, Sct, Cck-1r, Sctr, and Ctrb1 were designed by the software of Invitrogen™ Custom Oligonucleotides and are shown in Table 1. PCR amplification and quantification were carried out using an Eppendorf MasterCyclers ep RealPlex4 (Wesseling-Berzdorf, Hamburgers, Germany), as described previously [12] . The data were normalized to the amount of β-actin mRNA.