Fast start dna master sybr green 1 kit

Total RNA was extracted from the placentas using Trizol (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. RNA (1 μg) was used for reverse transcription using high-capacity cDNA of the reverse transcription kit RT (TaKaRa Biotechnology CO., Ltd) according to the instructions. The primer sequences of SULT1E1 and GAPDH mRNA were designed according to earlier publications34 (link). RT-qPCR was performed using a Light Cycler Fast Start DNA Master SYBR Green I kit and an ABI Prism 7300 Sequence Detection System (Applied Biosystems, Foster City, California, USA), and the relative expression of genes was determined using the 2-ΔΔct method with normalization to GAPDH expression. The results were averaged from four sets of independent experiments.

Midbrain containing substantia nigra was taken quickly, and total RNA was isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). RNA (1 μg) was used to reverse transcribe using high-capacity cDNA of the reverse transcription kit RT (TaKaRa Biotechnology CO., Ltd, Dalian, China). The primer sequences of DAT (forward primer: 5′-ATCAACCCACCGCAGACACCAGT-3′ reverse primer: 5′-GGCATCCCGGCAATAACCAT-3′) and VAMT2 mRNA (forward primer: 5′-ATGCTATCGGTCCCTCTGCTGGTG-3′; reverse primer: 5′-GACGGGGTACGGCTGGACATTATT-3′) were designed according to a previous publication.^{51 (link)} Q-RT-PCR was performed using a Light Cycler Fast Start DNA Master SYBR Green I kit and an ABI Prism 7300 Sequence Detection System (Applied Biosystems, Foster City, CA, USA). The relative expression of genes was determined using the 2-ΔΔct method, with normalization to GAPDH expression. The levels of DAT and VAMT2 mRNA were expressed as percent of WT mice.

The samples of the SN were quickly taken, and total RNA was isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). RNA (1 μg) was used to reverse transcribe using high-capacity cDNA of the reverse transcription kit RT (TaKaRa Biotechnology Co., Ltd.). The primer sequences of PPARγ (forward primer: 5′-GCTTATTTATGATAGGTGTGATC-3′; reverse primer: 5′-GCATTGTGAGACATCCCCAC-3′) and GAPDH mRNA (forward primer: 5′-ACCACAGTCCATGCCATCAC-3′; reverse primer: 5′-ACCACAGTCCATGCCATCAC-3′) were designed according to a previous publication^{11 (link)}. Q-RT-PCR was performed using a Light Cycler Fast Start DNA Master SYBR Green I kit and an ABI Prism 7300 Sequence Detection System (Applied Biosystems, Foster City, California, USA). The relative expression of genes was determined using the 2-ΔΔct method, with normalization to GAPDH expression. The levels of PPARγ mRNA were expressed as percent of control mice.