Practical Tips for Reliable cDNA Synthesis in qPCR Workflows
Why cDNA synthesis quality matters for qPCR results
In any qPCR workflow, the quality of your cDNA is critical. Poor reverse transcription can lead to low sensitivity, biased results, and unreliable gene expression data. By following a few best practices and using a high-performance kit such as the SensiFAST™ cDNA Synthesis Kit from Meridian Bioscience (Bioline), together with Meridian’s RNA Isolation Kits, researchers in Australia and New Zealand can achieve consistent, reproducible results from sample to data.
Tip 1: Start with High-Quality RNA
The integrity of your RNA will directly impact your cDNA synthesis. Meridian’s RNA Isolation Kits provide high-yield, inhibitor-free RNA suitable for sensitive downstream applications. Whether you are working with blood, tissues, or cultured cells, these kits deliver clean RNA that sets the foundation for reliable cDNA synthesis.
Tip 2: Choose a Kit Designed for Full Coverage
Not all cDNA synthesis kits are created equal. Some introduce a 3′ bias, leaving long transcripts under-represented. The SensiFAST cDNA Synthesis Kit uses a combination of anchored oligo(dT) primers (which ensure the 3′ ends of mRNAs are always captured) and random hexamer primers (which prime across the whole transcript). This ensures a more accurate reflection of gene expression.
Tip 3: Optimise for Low Input Samples
When working with small or precious samples such as biopsies, sensitivity matters. The SensiFAST kit can generate high-quality cDNA from as little as 1 picogram of RNA, making it ideal for low-input applications. Pairing it with Meridian’s RNA Isolation Kits ensures even small amounts of RNA are recovered efficiently and protected from inhibitors.
Tip 4: Validate with a Trial Kit
For labs trialling Bioline solutions for the first time, trial kits are available. These smaller packs let you test both the RNA Isolation Kit and the SensiFAST cDNA Synthesis Kit in your own workflows before scaling up—helping you validate results with minimal upfront cost.
Why Choose Millennium Science
As the exclusive distributor of Meridian Bioscience Bioline products in Australia and New Zealand, Millennium Science provides:
- Local stock availability for faster delivery
- Expert technical support across RNA isolation and cDNA synthesis workflows
- Competitive pricing and access to trial kits for first-time users
From RNA isolation to first-strand synthesis, every step matters in your qPCR workflow. By combining Meridian’s RNA Isolation Kits with the SensiFAST cDNA Synthesis Kit, researchers can achieve reproducible, high-quality results, even with challenging or low-input samples.