Quantitative Analysis of Hippocampal Genes

Hippocampal total RNA of mice was extracted using RNazol according to the manufacturer’s manuals (Takara, Dalian, China). After removal of the trace amounts of DNA contamination with DNase I, total RNA was reverse transcripted into cDNA with kit from Life Technologies (Grand Island, NY, USA). Quantitative PCR was carried out using Taqman SYBR kit (Life Technologies). The concentrations of target genes in the samples were determined by standard curves generated with template plasmids including fragments of the related target genes. At last, they were normalized to that of glyceraldehydes-3-phosphate dehydrogenase (GAPDH) in the same sample. The sequences for all the primers used were described as previously [25 (link)].

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He Y.X., Du M., Shi H.L., Huang F., Liu H.S., Wu H., Zhang B.B., Dou W., Wu X.J, & Wang Z.T. (2014). Astragalosides from Radix Astragali benefits experimental autoimmune encephalomyelitis in C57BL /6 mice at multiple levels. BMC Complementary and Alternative Medicine, 14, 313.

Publication 2014

RNazol Taqman SYBR kit

Cdna Dehydrogenase Dna contamination Dnase i Genes Mice Phosphate Plasmids Primers

Corresponding Organization :

Other organizations : Shanghai University of Traditional Chinese Medicine, China Pharmaceutical University, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Concentrations of target genes

control variables

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression
Removal of trace amounts of DNA contamination with DNase I

controls

Positive control: Template plasmids including fragments of the related target genes, used to generate standard curves
Negative control: Not explicitly mentioned

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