Unlock Reliable Gene Expression Analysis with Precision qPCR Assay Controls
Ensure accuracy and reproducibility in your gene expression studies with the proper use of qPCR assay controls. Our guide delves into the essential assays needed for effective qPCR and provides a comprehensive overview of their design and application.
Why Assay Controls Matter
Quantitative PCR (qPCR) is a powerful tool for studying gene expression, but the quality of your results depends heavily on the correct application of assay controls. From detecting genomic DNA contamination to evaluating RNA integrity and reverse transcription efficiency, each control step contributes to the accuracy of your results.
Types of Essential Assay Controls
- DNA Contamination Control Assay
Detects genomic DNA contamination in samples to ensure clean and reliable PCR results. - Positive PCR Control Assay
Validates PCR performance across samples, ensuring the reactions are working as expected. - RNA Quality Assay
Evaluates the integrity of RNA to determine if degradation may impact the PCR analysis. - Reverse Transcription Control Assay
Monitors the efficiency of reverse transcription, ensuring accurate conversion of RNA to cDNA. - Reference Gene Validation
Select and validate stable reference genes to normalize qPCR data, improving quantification accuracy. Bio-Rad offers prevalidated reference gene arrays for easy and reliable selection.
Step-by-Step Process to Ensure Accurate Results
1. Select Candidate Reference Genes
Choose from a list of well-known genes to test across your sample groups.
2. Isolate RNA & Reverse Transcription
Carefully prepare RNA from samples, ensuring consistent protocols for all conditions.
3. Perform qPCR & Analyze Stability
Test for gene expression stability and use the CFX Maestro Software to choose the most reliable reference genes.
4. Choose Stable Reference Genes for Normalization
Use ideal, stable genes for normalization to account for experimental variability and improve result accuracy.
Why Choosing the Right Controls is Crucial
Poor control selection can introduce significant errors in gene expression analysis, especially when studying subtle expression differences. The wrong reference genes—like common housekeeping genes (e.g., GAPD, ACTB)—can lead to incorrect conclusions due to their variability across conditions.
Tools and Resources
- PrimePCR Assays: Access a full range of prevalidated assays for DNA contamination, positive controls, RNA quality, and reverse transcription.
- CFX Maestro Software: Use advanced tools for analyzing reference gene stability and ensuring accurate normalization.
- Preplated 96/384-Well Reference Panels: Streamline your reference gene selection with Bio-Rad’s preplated arrays.
Get Started with PrimePCR
Make your gene expression analysis more reliable by integrating these essential controls into your qPCR experiments. Explore Bio-Rad’s comprehensive product range, including PCR primers, preplated panels, and advanced software, all designed to enhance the accuracy and reproducibility of your data.
This article is posted at bio-rad.com
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