RNA was isolated from sorted PB or SF myeloid subsets using RNeasy Micro Kit (Qiagen) according to manufacturer instructions. cDNA was synthesized using the reverse transcription kit (Promega), and gene expression was analyzed by semiquantitative PCR using the SYBR Green assay GoTaq qPCR Master Mix (Promega) with standardized primers (Metabion) on a StepOne Real-Time PCR system (Applied Biosystems). Relative gene expression was normalized to β-actin detection.
Sybr green assay gotaq qpcr master mix
Manufactured by Promega
The SYBR Green assay GoTaq qPCR Master Mix is a ready-to-use solution for quantitative real-time PCR (qPCR) analysis. It contains SYBR Green I dye, which binds to double-stranded DNA and emits fluorescence upon binding, enabling the monitoring of DNA amplification during the PCR process.
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Lab products found in correlation
2 protocols using sybr green assay gotaq qpcr master mix
1
Semiquantitative PCR of Myeloid Subsets
2
Cytokine Expression in Dendritic Cells
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RNA was isolated from cDC cultured in the absence or the presence of TBK1 adjuvants using RNeasy Micro Kit (Qiagen) according to manufacturer instructions. Subsequently, cDNA was synthesized using the Reverse Transcription kit (Promega). Transcriptional levels of IFN-, IL-12 and TNF were analyzed by semiquantitative PCR (SYBR Green assay Go Taq qPCR Master Mix; Promega) with specific primers (Metabion) on an Applied Biosystems StepOne Real-Time PCR system (Applied Biosystems). Relative gene expression was calculated after normalization to -actin transcriptional levels.
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