Device built-in

As a global leader in biotechnology, Roche's real-time quantitative PCR systems (qPCR) play a vital role in molecular biology research, clinical diagnostics, and environmental monitoring. Roche qPCR instruments are recognized as the gold standard for nucleic acid quantification, featuring high-precision temperature control, sensitive optical detection systems, and intelligent data analysis software. Below is a comprehensive introduction covering core technologies, product series, and application areas.

I. Core Technical Principles

Roche qPCR systems are based on real-time fluorescent quantitative PCR technology, enabling accurate quantification of initial templates by monitoring the accumulation of fluorescent signals during PCR amplification. Key technical features include:

• High-Precision Temperature Control System: Utilizes semiconductor Peltier technology with temperature control accuracy of ±0.1°C, well-to-well temperature variation ≤0.3°C, and maximum heating/cooling rates of up to 4.8°C/s, ensuring consistent amplification efficiency.

• Multi-Channel Fluorescence Detection: Equipped with 4-6 fluorescent detection channels (e.g., FAM, HEX, ROX, Cy5), supporting various detection systems such as SYBR Green dye method and TaqMan probe method, enabling multiplex PCR reactions.

• Melting Curve Analysis: Distinguishes specific amplification from non-specific products (e.g., primer dimers) based on differences in product melting temperature (Tm), verifying experimental specificity.

II. Main Product Series and Features

Roche's qPCR product line covers different throughputs and application needs, primarily including the following series:

1. LightCycler® Series

• LightCycler® 480: Medium-to-high throughput model supporting 96-well and 384-well plates (with optional modules), 6-color fluorescence detection, suitable for research and clinical batch testing. Its optical system uses a CCD camera to simultaneously detect all samples without optical edge effects, achieving single-copy level sensitivity.

• LightCycler® 96: Simplified model with a fixed 96-well plate format, offering high cost-effectiveness for routine laboratories. Equipped with four-color LED light sources and PMT detectors, supporting dyes such as FAM, HEX/VIC, ROX, and Cy5, with a dynamic range of 6-8 orders of magnitude.

2. cobas® Series

Models optimized for clinical diagnostics, such as the cobas® z 480, featuring fully enclosed detection systems and automated sample tracking functions. Compatible with FDA/CE-certified reagents (e.g., for HPV and HIV testing) and compliant with GLP/GMP standards.

III. Key Performance Advantages

1. Detection Sensitivity and Accuracy: Capable of detecting nucleic acids as low as 0.1 copy/μL, with a linear range spanning 10^1-10^9 copies, standard curve R² ≥ 0.995, repeatability CV < 0.15%, and the ability to distinguish concentration differences between 1000 and 2000 copies.

2. Efficient Operation: Completes 40 PCR cycles in as fast as 40 minutes (for 384-well plates), supporting gradient PCR and pre-cooling functions (4°C) to enhance experimental efficiency.

3. Intelligent Software: The accompanying LightCycler® software provides a guided operation interface, supporting absolute quantification (standard curve method), relative quantification (ΔΔCt method), melting curve analysis, SNP genotyping, and other functions. Data can be exported in various formats, including Excel and PDF.

IV. Typical Application Areas

1. Research Fields:

   • Gene expression analysis: Quantitatively detecting mRNA transcription levels to study gene regulation mechanisms.

   • CRISPR editing validation: Assessing editing efficiency by amplifying edited sites.

   • Microbial community research: Quantifying specific microbial populations in environmental or gut samples.

2. Clinical Diagnostics:

   • Viral load detection: Quantitative monitoring of viruses such as HBV, HCV, and HIV.

   • Tumor gene screening: Detecting mutations in cancer-related genes like EGFR and KRAS.

   • Genetic disease diagnosis: Mutation analysis for monogenic diseases such as cystic fibrosis and thalassemia.

3. Agriculture and Food Safety:

   • GMO component detection: Quantitative analysis of transgenic elements such as the 35S promoter and NOS terminator.

   • Foodborne pathogen monitoring: Rapid detection of Salmonella, Listeria, and other pathogens.

V. Supporting Products and Maintenance

Roche provides complete supporting reagents (e.g., FastStart TaqMan Probe Master Mix) and consumables (low-fluorescence background microplates). Original reagents are recommended to ensure stable performance. Instrument maintenance requires regular temperature calibration (every 6 months) and optical window cleaning. Daily operating conditions should be maintained at 15-30°C and 20-80% humidity.

VI. Technology Development Trends

With the growing demand for precision medicine and automation, Roche qPCR systems are evolving toward higher throughput (e.g., 384-well plates), faster detection (completing amplification within 30 minutes), and greater intelligence (LIMS system integration). Second-hand equipment (e.g., LightCycler® 96) remains practical for budget-limited laboratories but should be purchased through正规 channels with performance verification.

Roche qPCR systems continue to provide accurate and efficient nucleic acid quantification solutions for life science research and molecular diagnostics, leveraging their reliable technology platform and extensive application validation.