Vazyme - For Science For Health

Gene Expression Analysis

Gene expression analysis is a cornerstone of modern biotechnology, enabling the precise measurement of transcriptional products like mRNA in cells, tissues, or entire organisms. This powerful technique provides critical insights into gene activity and function, shedding light on how genes respond to varying environmental conditions and physiological states. Such studies play a pivotal role in unraveling complex biological processes and advancing our understanding of disease mechanisms.

Core Challenges We All Face

However, maximizing the potential of gene expression analysis is not without its challenges. Are any of the following issues familiar to you?

RNA sample quality issues: Sample degradation, insufficient RNA purity, or limited sample quantity can lead to inaccurate downstream results.
Complex experimental procedures: Tedious reverse transcription and qPCR steps, involving multiple reagents and pipetting steps, increase the likelihood of errors and reduce experiment reproducibility.
Non-specific amplification: Non-specific peaks or primer-dimers in the melting curve interfere with accurate detection of target genes.
Insufficient detection sensitivity: Low-expressing genes or rare targets are difficult to detect reliably, resulting in high C_T values.
Reagent compatibility issues: qPCR reagents may have poor compatibility with different instruments, requiring manual addition or adjustment of ROX reference dye.
Challenges with standardization: Difficulty in selecting reference genes and lack of uniform experimental conditions undermine data reliability.
High risk of contamination: Genomic DNA (gDNA) contamination affects the specificity and accuracy of quantitative results.

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Vazyme Gene Expression Analysis Solution

Proud Winner of the 2024 Bioz Top Star Award in qPCR Assays

We understand the critical importance of precision and reproducibility in gene expression studies. This commitment has earned global recognition, as our qPCR assays were named the 2024 Bioz Top Star™ Award Winner in the qPCR Assays category—a distinction based on AI-powered analysis of over 36 million scientific publications.

This accolade underscores our commitment to delivering tools trusted by researchers worldwide to overcome today’s most pressing challenges in gene expression analysis. With 15,000+ researchers globally relying on our technology, and 7.8 million reaction units deployed in 2024 alone, this accolade underscores our ongoing efforts to delivering tools trusted by scientists worldwide to overcome today’s most pressing challenges in gene expression analysis.

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Vazyme Workflow

As a professional supplier of products and solutions in the life sciences and in vitro diagnostics fields, Vazyme has accumulated years of insight and research into scientific needs.

To address these challenges, we offer a comprehensive solution that covers every step of the workflow—from RNA extraction to reverse transcription and qPCR. Our product lineup includes reagents, consumables, and instruments, providing a seamless experience for researchers. Designed with simplicity and efficiency in mind, our solutions are user-friendly, making it easy even for rookies to successfully complete the entire experiment. With our professional and fully integrated system, you can achieve more reliable and accurate results, paving the way for groundbreaking discoveries and high-impact publications.

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Vazyme Comprehensive Solutions

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Introducing our Rapid & Easy-to-Use RNA Extraction Kit—designed to streamline your research with both Cell/Tissue and Plant-specific solutions. Each kit is engineered for high efficiency, providing a quick and safe way to extract RNA with minimal effort. Our kits come equipped with a gDNA removal column to ensure high-quality, contaminant-free RNA, enabling reliable results in downstream applications. Whether you're working with animal cells, tissues, or plant samples, we have the perfect solution to meet your needs and accelerate your research.

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Introducing our qPCR cDNA Synthesis Kit—engineered to support your research with both Classic and Upgraded Series. Each series offers two options: SuperMix for superior RT efficiency and All-in-One for unmatched speed and convenience. The Classic Series provides consistent, reliable performance for standard applications, while the Upgraded Series is tailored for challenging samples, including those with inhibitors, complex secondary structures, or degraded RNA—ideal for FFPE and low-quality samples. Whatever your sample type, we have the optimal solution to simplify and enhance your workflow.

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Superior Reverse Transcription Efficiency

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100 ng RNA from animals, plants and microbes were reverse transcribed by Vazyme #R423 and similar RT products from Suppliers. The RT efficiency of these reagents was evaluated by qPCR amplification of obtained cDNA. It can be seen from the figure, ΔC_T between Vazyme #R423 and Supplier A/B are more than 0.5, indicating a better efficiency of Vazyme #R423.

Excellent Template Compatibility

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100 ng human FFPE samples and degraded 293T cells (Rin=1) were reverse transcribed by Vazyme #R423 and similar RT products from Suppliers. The template compatibility of these reagents was evaluated by qPCR amplification of obtained cDNA. The results show that ΔCT between Vazyme #R423 and Suppliers are more than 0.5 in most genes, indicating a better template compatibility of Vazyme #R423.

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Introducing our qPCR MasterMix—available in both Classic and Next-Gen Series to meet your research needs. The Classic Series ensures stable detection of low-expression systems and includes universal ROX for effortless compatibility with all qPCR instruments. The Next-Gen Series builds upon the Classic with enhanced features for low-abundance targets, complex templates, and contamination-free experiments—perfect for demanding applications. No matter your research challenge, we provide the right solution to streamline your workflow.

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Great Balance Between Sensitivity and Specificity

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Fig. A shows eight 10-fold gradient dilutions of pUC19 plasmid and Fig. B shows the standard curve obtained according to the C_T values in Fig. A. The plasmid copy number concentration in the eighth gradient was about 6 copies/4 µl. pUC19 in each dilution gradient was assayed with Vazyme #Q712, and the template volume in each well was 4 µl. The results show that Vazyme #Q712 can determin the template copy number with the high sensitivity. In Fig. C, cDNA of HeLa cells was used as a template, six different genes were amplified under the same reaction conditions using Vazyme #Q712 and SYBR qPCR reagents of other suppliers. The results show that Vazyme #Q712 can detect target genes with a great specificity.

All Platform Compatibility

$\"\"/$ Vazyme #Q712 was used to amplify EGFR gene on different qPCR instruments (Type: ABI Stepone, ABI QuantStudio 3, and Roche LightCycler 480) . The results show the wide applicability of Vazyme #Q712 for instruments and the concentration of ROX does not need to be adjusted on different instruments.

Frequently Asked Questions

Q1: What's the working principle of SYBR Green?

Definition: Dye-based qPCR uses fluorescent dyes (such as SYBR Green) to detect amplified DNA during the PCR process. As the DNA is amplified, these dyes bind to double-stranded DNA, emitting a fluorescence signal that is proportional to the amount of DNA produced.

Q2: What should you pay attention to when designing primers for qPCR?

(1)Amplicon Length: The recommended length of the amplification product is 80-200 bp： Shorter products improve amplification efficiency. The minimum length should be greater than 80 bp to differentiate from primer-dimers effectively. Overly long amplicons reduce efficiency, so striking this balance is critical for achieving a 90% success rate. (2)GC or AT Content: Avoid regions with high GC or AT content, especially at the 3′ end of the primer. This minimizes the risk of secondary structures or nonspecific binding. (3)3′ Base Preference: The terminal base at the 3′ end should ideally be G or C, as these bases provide stronger binding stability. Avoid ending with T. (4)Primer Pair Tm Values: Ensure that the melting temperatures (Tm values) of the forward and reverse primers differ by no more than 1°C to maintain consistent annealing conditions. (5)GC Content: Aim for a GC content of 40-60% to ensure primer stability and specificity. (6)For Probe-Based Assays: If using a probe, its Tm value should be 8-10°C higher than the Tm value of the primers to optimize hybridization specificity.

Q3: Should cDNA Templates Be Diluted? How Much Should They Be Diluted for Quantification?

The dilution factor for cDNA templates depends on your experiment and there isn’t a universal standard. Generally, for every 10-fold dilution of cDNA, the Ct value increases by approximately 3.3 cycles. You can use this relationship to determine the optimal dilution for your samples. (1)Perform Preliminary Dilution Tests: Use the cDNA stock solution, 10-fold dilution, and 100-fold dilution as templates in preliminary qPCR runs. Select the dilution factor that results in a Ct value between 18-28, as this range ensures accurate quantification. (2)Consider Previous Conditions: If switching reagents or systems, start with the same dilution factor you used previously, and adjust as needed based on results. (3)Avoid High Concentration Risks: When using undiluted cDNA stock as the template: -Do not exceed 1/10 of the total reaction volume, as undiluted cDNA often contains qPCR inhibitors. -Using excessive volumes increases the risk of nonspecific amplification or poor amplification efficiency. By carefully diluting and optimizing cDNA input, you can minimize interference and achieve reliable quantification results.