Quantitect syrb green dye

Gene expression study for the genes of our interest was measured by real-time quantitative RT-PCR (Step one Applied Biosystems) using QuantiTect SYRB Green dye (Qiagen, Valencia, CA).
All the primer sets were provided by Qiagen (npy: Dr_npy_1_SG QuantiTect Primer Assay (QT02205763).
HMGB1: Dr_hmgb1b_2_SG QuantiTect Primer Assay (Cat no: QT02088555)
TLR4: Dr_tlr4ba_va. 1_SG QuantiTect Primer Assay (Cat no: QT02198539)
NF-κB: Dr_nfkb1_2_SG QuantiTect Primer Assay (Cat no: QT02498762)
TNF-α: Dr_tnf_1_SG QuantiTect Primer Assay (Cat no: QT02097655)
IL-1: Dr_il1rapl1a_1_SG QuantiTect Primer Assay (Cat no: QT02131850)
BDNF: Dr_bdnf_1_SG QuantiTect Primer Assay (Cat no.QT02125326)
CREB_1: Dr_CREB_1 bpa_1_SG QuantiTect Primer Assay (Cat no. QT02197503)
NPY: Dr_npy_1_SG QuantiTect Primer Assay (Cat no: QT02205763)
eef1a1b: Dr_eef1a1b_2_SG QuantiTect Primer Assay (Cat no: QT02042684)
The PCR mixture contained 1X SYBR green PCR master mix (Qiagen), 0.7 µM each forward and reverse primers, and 1 µL of sample cDNA. Samples were incubated at 95 °C for 2 min before thermal cycling (40 cycles of 95 °C for 5 s and 60 °C for 15 s). Relative expression values of the above genes were obtained by normalizing threshold cycle (Ct) values of genes of interest against Ct value of eef1a1b (housekeeping gene) (2^^{[Ct eef1a1b-Ct Gene of interest]}).

Gene expression for NPY, BDNF and CREB1 were computed using real-time quantitative RT-PCR (Applied Biosystems) using QuantiTect SYRB Green dye (Qiagen, Valencia, CA, United States). All the primer sets were purchased by Qiagen (npy: Dr_npy_1_SG QuantiTect Primer Assay (QT02205763), bdnf: Dr_bdnf_1_SG QuantiTect Primer Assay (QT02125326), creb:Dr_crebbpa_1_SG QuantiTect Primer Assay (QT02197503). Samples were incubated at 95°C for 2 min prior to thermal cycling (40 cycles of 95°C for 5 s and 60°C for 15 s). Relative expression of these three genes were attained by normalizing threshold cycle (Ct) values against Ct value of eef1a1b (housekeeping gene) (2 ^∧ [Ct eef1a1b – Ct Gene of interest]).