Episode 3: qPCR in Molecular Research – Best Practices, Pitfalls and Lessons Learned
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Quantitative PCR (qPCR) is a foundational tool in molecular research, delivering precise and sensitive gene expression measurements. However, ensuring data reliability and reproducibility demands strict adherence to methodological best practices. In the final episode of our New Frontiers in PCR Technology series, we will explore the complexities of qPCR and examine critical lessons learned from past misuse.
Key topics will include:
- The development of the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines to standardise qPCR methodology
- Challenges posed by reverse transcription efficiency and the proper selection and validation of reference genes for accurate data normalization
- Common analytical errors, such as the inappropriate use of the ΔΔCq method without considering PCR efficiency
- Underestimation of error propagation in published qPCR studies
- Actionable recommendations for enhancing methodological rigour in qPCR
Attendees will come away with actionable insights to improve their qPCR experiments, from adhering to MIQE guidelines to optimizing replicates and employing robust data analysis techniques. This episode will offer practical recommendations for enhancing methodological rigor and ensuring ongoing accuracy in molecular research.