Breaking Through RT-qPCR boundaries in Gene Expression Analysis

Variation in gene expression among cell types, tissues, and organisms is commonly examined by reverse transcription quantitative PCR, or RT-qPCR. In this process, RNA is isolated from samples of interest and reverse-transcribed into complementary DNA (cDNA). The levels of gene expression are then determined from the quantity of cDNA amplified by PCR. The initial stages of the RT-qPCR workflow, such as RNA isolation and reverse transcription, can critically impact the accuracy of the quantitation of gene expression.

Learning Objectives:
1. Discuss important considerations for RNA isolation.
2. Review tools for efficient cDNA preparation.
3. Clarify sources of poor RT-qPCR results.