Quantitative PCR (qPCR) and reverse transcription qPCR (RT-qPCR) are indispensable tools across life sciences, diagnostics, and biotech. But in practice, traditional workflows are often a bottleneck—especially in POCT (Point-of-Care Testing) environments where speed, simplicity, and reproducibility are critical.
Fortunately, fully premixed, one-tube qPCR solutions are transforming molecular detection workflows—making them faster, more consistent, and highly compatible with portable, automated qPCR platforms used in decentralized settings.
Figure 1. Standard workflow VS Yeasen Full Premix workflow
The Challenges: Traditional qPCR Is Cumbersome
In traditional lab workflows, qPCR setup is time-consuming and error-prone:
- Multiple pipetting steps (enzymes, primers, probes, buffers)
- Higher contamination risk
- Low throughput reproducibility
- Slow turnaround (setup alone can take 30–60 minutes)
In a POCT context—such as field surveillance, urgent care clinics, or mobile labs—these problems are magnified. There's little room for human error, and every extra step slows down result delivery.
The Solution: Fast, Fully Premixed, One-Tube qPCR Systems Built for POCT
One-tube qPCR mixes consolidate all reaction components into a single master mix(Supports pre-mixing of primers and probes for multiplex applications)—optimized for fast setup, robustness, and instrument compatibility, including compact qPCR platforms used at the point of care.
Key Advantages:
- Ultra-fast setup – Mix-and-run in under 20 minutes, even for large batches.
- Excellent premix stability: Stable after 7 days at 37°C and 20 freeze-thaw cycles.
- Low-DNA residue: No detectable residual DNA from E. coli, P. aeruginosa, or human genomic DNA across 48 wells.
- Fewer errors, less training – Simplified protocols minimize the need for skilled personnel.
Products Properties
|
Category |
Performance Specifications |
|
Specificity |
(1) At time T0, 16 wells across multiple targets show no amplification, indicating high specificity. (2) After accelerated aging at 37°C for 7 days in a fully premixed format, show no amplification, indicating improved specificity. |
|
Linear Range |
Meets the required range: 10–10⁷ copies/μL, with R² ≥ 0.98 and amplification efficiency of 100% ± 10%. |
|
Precision |
Across high, medium, and low template concentrations (CT values from 21 to 37), the coefficient of variation (CV) is less than 2%. |
|
Fast Protocol Compatibility |
Compatible with the fast program of the SLAN-48S instrument (20 minutes). Compared to the standard 50-minute program, the CT value difference for targets is within 0.6, with no loss in detection sensitivity. |
|
Stability |
(1) No change in amplification performance after 20 freeze-thaw cycles compared to −20°C storage. (2) Stable amplification performance after 7 days at 37°C (reagent itself) and after 7 days at 4°C. (3) Supports full premix storage at 37°C for 7 days with unchanged performance. |
|
Sensitivity |
Detects as low as 0.125 copies/μL, equivalent to single-copy detection per reaction. |
1. Linear Range
Figure 2. Amplification efficiency test of N. meningitidis and S. pneumoniae using fast cycling protocols.
(A) ABI 7500 Fast Mode: N. meningitidis (R² = 0.999, Eff = 99.864%), S. pneumoniae (R² = 1.000, Eff = 98.952%)
(B) ABI Q5 Fast Mode: N. meningitidis (R² = 0.999, Eff = 99.14%), S. pneumoniae (R² = 1.000, Eff = 99.756%)
2. Precision
Figure 3. qPCR precision analysis using three different template concentrations. The assay showed high precision with consistent CT values and low coefficient of variation (CV < 2%) across all levels.
3. Fast Protocol Compatibility
|
Instrument |
SLAN |
System |
5 μL DNA(25 μL total volume) |
|
Template |
bacterial plasmid template (B-tube and P-tube). |
Template concentrations |
Ct 30-35 |
|
Progarm |
Conventional program (96S/48S): 95°C 5 min；40 cycles (95°C 15sec；60°C 30 sec)--50 min Fast program (48S Fast): 95°C 15 sec；40 cycles (95°C 2sec；60°C 8 sec)--20 min |
||
Table 1. Sensitivity Evaluation of 16924 Under Fast and Conventional Programs
|
16924 |
|
Group B |
Group P |
|||||
|
96S-Conventional program |
|
FAM |
VIC |
ROX |
FAM |
VIC |
CY5 |
ROX |
|
Mean Ct Value |
36.16 |
36.13 |
33.23 |
35.61 |
38.81 |
NA |
36.03 |
|
|
Detection Rate |
100%(16/16) |
100%(16/16) |
100%(16/16) |
75%(12/16) |
81.25%(13/16) |
0 |
93.75%(15/16) |
|
|
48S-Fast program |
|
FAM |
VIC |
ROX |
FAM |
VIC |
CY5 |
ROX |
|
Mean Ct Value |
36.54 |
36.31 |
33.54 |
36.46 |
39.14 |
36.87 |
36.31 |
|
|
Detection Rate |
100%(16/16) |
100%(16/16) |
100%(16/16) |
87.5%(12/16) |
68.75%(11/16) |
12.5%(2/16) |
81.25%(13/16) |
|
|
48S-Fast program |
|
FAM |
VIC |
ROX |
FAM |
VIC |
CY5 |
ROX |
|
Mean Ct Value |
36.30 |
36.34 |
33.77 |
35.54 |
38.73 |
39.41 |
35.58 |
|
|
Detection Rate |
100%(16/16) |
100%(16/16) |
100%(16/16) |
87.5%(14/16) |
56.25%(9/16) |
6.25%(1/16) |
87.25%(14/16) |
|
Figure 4. Performance comparison of 16924 and Supplier A* using conventional and fast programs.
Two sets of 4-plex plasmid templates (B-tube and P-tube) were tested using both 16924 and Supplier A* at relatively high template concentrations (Ct 30–35). Supplier A* showed over 1 cycle delay in Ct values under the fast program for P-tube templates, while 16924 showed no significant difference between programs. Some targets in 16924 amplified earlier in the fast protocol.
4. Strong stability
Figure 5. Stability evaluation of the qPCR Mix under various conditions. The mix showed consistent amplification performance after long-term storage at −20°C, 20 freeze–thaw cycles, and accelerated aging at 37°C for 7 days, demonstrating excellent stability.
5. High inhibitor tolerance
|
Instrument |
SLAN 96S |
System |
5 μL DNA(25 μL total volume) |
|
Template |
bacterial plasmid template (Different dilutions). |
Template concentrations |
Ct 32-38 |
|
Progarm |
95°C 5 min；45 cycles (95°C 15sec；60°C 30 sec) |
||
Figure 6. Comparative performance of 16924 and a commercial product (Supplier A) using 4-plex plasmid templates diluted in different buffers.
Figure 7. Comparison of qPCR performance between 16924 and Supplier A using different diluents
Templates diluted in various solutions, including TE buffer and saline, were tested with 16924 and Supplier A*. The results show that 16924 exhibits greater tolerance to diverse dilution buffers—particularly saline—providing more reliable detection in buffer-restricted or field-based settings.
6. Sensitivity
Figure 8. Sensitivity and stability evaluation of the qPCR Mix. A plasmid template was serially diluted from 10 copies/μL to 0.125 copies/μL and tested in 8 replicate wells per concentration. The mix maintained consistent detection performance after storage at −20°C, 20 freeze–thaw cycles, and accelerated aging at 37°C for 7 days.
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