Polymerase Chain Reaction (PCR) is the cornerstone of modern molecular biology, enabling researchers to amplify and analyse genetic material with exceptional precision. Quantitative PCR (qPCR) takes this capability further by measuring DNA or RNA levels in real time, allowing laboratories to obtain accurate, reproducible results for diagnostics, research, and quality control. Apex Scientific, a trusted laboratory equipment supplier in South Africa, provides advanced qPCR solutions and expert support to help scientists achieve consistent success.
Understanding PCR and qPCR
Conventional PCR amplifies DNA and reports results at the end of the process. In contrast, qPCR—also known as real-time PCR—monitors amplification during each cycle using fluorescent dyes or probes. This real-time measurement provides quantitative data on gene expression, pathogen detection, and genetic variation. Choosing the right qPCR instrument is therefore critical for laboratories seeking fast, reliable results.
Why qPCR Matters in Modern Laboratories
qPCR enables sensitive detection of nucleic acids for clinical diagnostics, food safety testing, environmental monitoring, and pharmaceutical research. South African laboratories increasingly rely on qPCR to track infectious diseases, monitor water quality, and evaluate agricultural samples. Accurate PCR data underpins important public health and research decisions, making dependable equipment and correct technique essential.
Key Technical Specifications to Consider
When selecting a qPCR system, evaluate these important factors:
- Optical system and detection chemistry – Ensure compatibility with SYBR®, TaqMan®, and multiplex assays.
- Throughput and capacity – Choose a format that matches your laboratory’s sample volume and growth plans.
- Temperature uniformity – Precise thermal control is critical for reproducible PCR results.
- Software and connectivity – Intuitive analysis tools and network capabilities simplify data sharing and compliance.
- Service and support – Reliable local assistance from an experienced equipment supplier ensures long-term value.
Top Products for qPCR
Apex Scientific proudly supplies the Azure Cielo Real-Time PCR System, designed for accuracy and ease of use. This instrument offers advanced optical technology, rapid cycling, and user-friendly software to streamline every qPCR workflow.
Explore detailed specifications on the Azure Biosystems Cielo page.
Five Tips for a Successful qPCR Run
1. Design Great Primers and Probes
- Determine a suitable reference gene; validated primers may already exist.
- Target an amplicon size of 70–150 bp and span an exon-exon junction.
- Maintain GC content between 40 % and 60 %, with melting temperatures of 50–65 °C.
- Avoid long nucleotide runs and secondary structures.
- Verify specificity using Primer-BLAST and test primer dimers with IDT’s OligoAnalyzer™.
- Optimise annealing temperature using a gradient run on your qPCR device.
- Store oligos at –20 °C in nuclease-free water.
2. Isolate High-quality RNA
- RNA is easily degraded; work on ice, wear PPE, and sterilise tools with H₂O₂.
- Confirm purity with a 260/280 ratio of 1.8–2.0 and assess integrity on a gel.
- Treat samples with DNase to remove genomic DNA before cDNA synthesis.
3. Refine Template Quantity and Reaction Efficiency
- Vortex and briefly spin down reagents before use.
- Perform serial dilutions to achieve ideal cycle threshold (Ct) values between 20 and 30.
- Calculate reaction efficiency using a standard curve and aim for R² > 0.95.
- Validate multiplex assays together to ensure consistent amplification.
4. Include the Proper Controls
- Use a no-template control to check for contamination.
- Include a positive control to confirm reaction conditions and primer performance.
- Run a no-reverse-transcriptase control to detect genomic DNA contamination.
- For SYBR® assays, add a melt curve to verify single-product amplification.
- Incorporate an internal positive control to detect PCR inhibitors.
5. Select the Appropriate Method for Ct Analysis
- Decide between threshold or regression analysis based on your experimental design.
- Set the baseline two cycles earlier than the Ct of the most abundant sample for threshold analysis.
- Confirm that cycling conditions match your chosen analysis method.
Essential Accessories and Options
Successful PCR experiments also depend on reliable accessories such as high-quality plasticware, calibration kits, and validated reagents. Apex Scientific provides consumables, calibration tools, and compatible software modules to keep your qPCR workflows consistent and reproducible.
Choosing the Right PCR Equipment for Your Laboratory
When evaluating different qPCR instruments, consider current and future sample volumes, budget constraints, and service requirements. Look for scalable systems with upgrade paths to accommodate growth. Laboratories in South Africa benefit from selecting an equipment supplier that offers local training, spare parts, and responsive technical support.
Conclusion
High-quality PCR data begins with careful planning, robust technique, and reliable equipment. By following the tips above and choosing a proven qPCR platform such as the Azure Cielo, your laboratory can achieve accurate, reproducible results.
Visit our Apex Scientific qPCR page to explore specifications, request a quote, or speak with our team about your application needs.
