Sybr green 2 master

Total tissue RNA was extracted by TRIzol reagent (Invitrogen) in accordance with the manufacturer's protocols and quantified by NanoDrop 2000 spectrophotometer, 1 µg RNA was used for reverse transcription with the Reverse Transcription System Kit (Promega) to cDNA. Real-time PCR amplification and detection was performed using the SYBR Green II Master (Takara) on LightCycler 480 (Roche Applied Science). The expression levels were normalized to housekeeping 36b4 gene. The primers used in this study are listed in (Supplementary Table 1, see section on supplementary data given at the end of this article).

Total RNA was extracted from tissues using TRIzol Reagent (Invitrogen), and 1 μg of RNA was transcribed following the manufacturer's instruction by using the Reverse Transcription System Kit (Promega), which produced high-quality complementary DNA. A real-time polymerase chain reaction was performed on LightCyclerH 480 (Roche) using SYBR Green II Master (Takara). All mRNA quantification data were normalized to the housekeeping 36B4 gene. All samples were run in duplicate in a single 384-well reaction plate, and data of the reaction were collected and analyzed according to the ΔΔC T method. Primers used in this study are listed in Supplementary Table 1, see section on supplementary data given at the end of this article.