How can we select housekeeping gene(s) for real-time PCR data analysis?

How can we select housekeeping gene(s) for real-time PCR data analysis? Selecting a (or more than one) proper housekeeping gene(s) is not an easy job. RT-qPCR is frequently chosen as the preferred method to observe gene expression because it offers sensitive and precise detection, the ability for high throughput, quick and precise quantification, and has a high potential for automation. However, to ensure that gene expression results are both precise and comparable among various experimental setups, conditions, operators, and laboratories, it is necessary to normalize RT-qPCR data against one or two housekeeping genes. These housekeeping genes must have a constant cellular expression, regardless of the experimental conditions, which is essential for obtaining reliable results. Additionally, when choosing an housekeeping gene for normalization of RT-qPCR data, it is important to consider the stability of its expression across various specimens and experimental conditions. This consideration should not be disregarded since the incorrect selection of an housekeeping gene can result in inaccurate data and misleading outcomes.

In the following articles, there are a few tips you can adopt to find proper housekeeping genes for yourself.

1. Identification of the best housekeeping gene for RT-qPCR analysis of human pancreatic organoids .
2. Importance of Housekeeping gene selection for accurate RT-qPCR in a wound healing model .