The relative gene expression including miR-181b, Arg-1, iNOS, CCR7, CD206, ALP, RUNX2, OCN, VEGF and BMP-2 was detected by qRT-PCR. Firstly, we applied TRIzol reagent (Invitrogen) for extracting the total RNA of RAW264.7 and hBM-MSC. Then, PrimeScript RT reagent Kit (Takara; Vazyme Biotech) was used for reverse transcription for acquiring complementary DNA of the extracted total RNA. Subsequently, qRT-PCR analysis experiments were conducted by SYBR Green detection reagent system (Takara). Finally, the levels of relative gene expression were calculated by applying the 2^−(△△CT) and 18S RNA levels was used for normalization. The primers sequence can be seen in Supplementary material.
Sybr green detection reagent system
Manufactured by Takara Bio
The SYBR Green detection reagent system is a real-time PCR reagent that utilizes the SYBR Green dye to detect and quantify target DNA sequences. The SYBR Green dye binds to double-stranded DNA, and the fluorescence signal produced is proportional to the amount of amplified DNA present in the reaction.
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Lab products found in correlation
2 protocols using sybr green detection reagent system
1
Relative Gene Expression Analysis
2
Multiplex Gene Expression Analysis
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The relative gene expression including miR-181b, Arg-1, iNOS, CCR7, CD206, ALP, RUNX2, OCN, VEGF and BMP-2 was detected by qRT-PCR. Firstly, we applied TRIzol reagent (Invitrogen) for extracting the total RNA of RAW264.7 and hBMSC. Then, PrimeScript RT reagent Kit (Takara; Vazyme Biotech) was used for reverse transcription for acquiring complementary DNA of the extracted total RNA. Subsequently, qRT-PCR analysis experiments were conducted by SYBR Green detection reagent system (Takara). Finally, the levels of relative gene expression were calculated by applying the 2^-(△△CT) and 18S RNA levels was used for normalization.
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