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This protocol outlines the steps for synthesizing complementary DNA (cDNA) from RNA samples, followed by quantitative PCR (qPCR) amplification to analyze gene expression. The process begins with RNA extraction from cultured cells, followed by cDNA synthesis using M-MLV reverse transcriptase. The resulting cDNA is then used as a template in qPCR, where gene expression is quantified using SYBR Green chemistry and specific primers. This method is widely used to investigate the expression levels of target genes in various biological contexts, such as response to treatments or genetic modifications.

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cDNA Synthesis and Quantitative PCR (qPCR) for Gene Expression Analysis

  • 实验方案

  • 发表日期：

    2025年

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  • Karuppusamy Arunachalam,
  • Puthanpura Sasidharan Sreeja

摘要

This protocol outlines the steps for synthesizing complementary DNA (cDNA) from RNA samples, followed by quantitative PCR (qPCR) amplification to analyze gene expression. The process begins with RNA extraction from cultured cells, followed by cDNA synthesis using M-MLV reverse transcriptase. The resulting cDNA is then used as a template in qPCR, where gene expression is quantified using SYBR Green chemistry and specific primers. This method is widely used to investigate the expression levels of target genes in various biological contexts, such as response to treatments or genetic modifications.