Taqman sybr kit

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)].

Total RNAs from the tissues or cells were extracted using Trizol reagent (Life Technologies). After eliminating trace amounts of DNA contamination with DNase I, total RNA was reversely transcripted into cDNA with Prime Script RT reagent (TakaRa, China). Quantitative PCR was carried out using Taqman SYBR kit (Life Technologies). Quantity of target genes was determined by standard curves generated with template plasmids containing fragments of respective target genes. Thereafter, they were normalized to that of glyceraldehydes-3-phosphate dehydrogenase (GAPDH) in the same sample. The sequences of forward and reverse primers were listed as follows: for iNOS, 5′-AACATCAGGTCGGCCATCAC-3′ and 5′-CCAGAGCCTGAAGTCATGTTTG-3′; for TNFα, 5′-AACCTCCTCTCTGCCGTCAAG-3′ and 5′-CCTCCCAGGTATATGGGCTCAT-3′; for IL1β, 5′-TGGGCCTCAAAGGAAAGAATC-3′ and 5′-GGTATTGCTTGGGATCCACACT-3′; for CD11b, 5′-GGTCGGCAAGCAACTGATTT-3′ and 5′-CAACTTGCATTATGGCATCCA-3′; for GAPDH, 5′-ATGTGTCCGTCGTGGATCTGA-3′ and 5′-ATGCCTGCTTCACCACCTTCT-3′.