Protocol detail

Quantitative PCR Analysis of RNA

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Total RNA from BV2 cells and Sp5 tissues were derived with TRIzol Reagent (Invitrogen, USA) and reversed to cDNAs using RevertAid Master Mix with DNase I (Thermo Fisher Scientific, USA). cDNA samples, gene specific primers (Table 1) and SYBR™ Green Master Mix (Thermo Fisher Scientific) were used to perform qPCR experiments according to the protocol, which were collected by ABI StepOnePlus PCR system (ABI-7500, Thermo Fisher Scientific). The relative mRNA expressions of genes were calculated by the 2^-△△CT method as described previously (18 (link)).

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Chen Y., Peng F., Yang C., Hou H., Xing Z., Chen J., Liu L., Peng C, & Li D. (2023). SIRT1 activation by 2,3,5,6-tetramethylpyrazine alleviates neuroinflammation via inhibiting M1 microglia polarization. Frontiers in Immunology, 14, 1206513.

Publication 2023

TRIzol Reagent RevertAid Master Mix with DNase I SYBR™ Green Master Mix ABI StepOnePlus PCR system

Cdna Cells Dnase i Gene Genes expressions Mrna Primers Sybr green Tissues Trizol

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Variable analysis

independent variables

RNA source (BV2 cells, Sp5 tissues)

dependent variables

Relative mRNA expressions of genes

control variables

TRIzol Reagent (Invitrogen, USA) for RNA extraction
RevertAid Master Mix with DNase I (Thermo Fisher Scientific, USA) for cDNA synthesis
Gene-specific primers (Table 1)
SYBR™ Green Master Mix (Thermo Fisher Scientific)
ABI StepOnePlus PCR system (ABI-7500, Thermo Fisher Scientific) for data collection
2^-△△CT method for data analysis

controls

No positive or negative controls were explicitly mentioned.

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