Fluorescent quantitative PCR is a highly sensitive experiment that requires a high degree of precision in the operation by laboratory personnel. Therefore, during the qPCR experiment process, we inevitably encounter problems:
I. Issues related to non-experimental factors
- The laboratory has many qPCR experiment projects, and the instruments are occupied, leaving the carefully prepared reaction mixtures with no clear disposal method.
- There are numerous quantitative samples, the task of adding samples is onerous, and there is a risk of sample addition errors due to dizziness, leading to unreliable experimental results.
- Long machine run times, limited number of experimental runs, and experimental tasks cannot be completed as planned on a regular basis.
- The laboratory equipment is being replaced with new instruments, there are multiple quantitative instruments in the laboratory, and the quantitative reagents are not compatible with the instruments.
II. Issues limited by experimental factors
Quantification issues with low-abundance genes. Complex primer design and screening processes, sensitivity, and reproducibility of replicate wells...
As a seasoned manufacturer dedicated to the research and development of qPCR reagents, YEASEN has meticulously developed a high-performance qPCR premix solution – Hieff UNICON™ Universal Blue qPCR SYBR Green Master Mix, which can solve the aforementioned experimental woes in one go. The Universal ROX used in this product replaces the traditional ROX, compensating for the optical path differences across various instrument platforms, making it suitable for all qPCR instruments. Additionally, the product employs an antibody modification method and includes additives that enhance gene amplification, significantly improving the amplification performance of the product. It perfectly balances the high-performance needs of researchers in the scientific community with the high-efficiency requirements of industrial enterprises.
Product Features
- Wide instrument compatibility: The premix contains a specific type of reference dye, eliminating the need to adjust the ROX concentration.
- Easy Tracing: The blue premix allows for direct visualization to distinguish whether the sample has been added, simply by observing the color.
- High Stability: The system is prepared at room temperature, and the mixed system can be stored at 4°C for up to 24 hours without any change in the test results.
- Rapid Setup: Compatible with both standard and fast programs, allowing the quantitative experiment to be completed in as fast as 46 minutes.
- Excellent Amplification Performance: High amplification efficiency, good specificity, capable of detecting genes at the single copy number.
Product Performance
1.High sensitivity: capable of detecting down to a single copy
Figure 1. Hieff UNICON™ Universal Blue premix can effectively detect a template amount range across 7 orders of magnitude, with high amplification efficiency and a good linear relationship within a broad linear range. The template used was 2 µL of plasmid with a concentration of 10^6 to 10^0 copies, amplifying the human IL23R gene.
2.The detection rate and specificity are good: widely applicable to amplimers with 30-70% GC content
Figure 2. Hieff UNICON™ Universal Blue premix is widely applicable to amplimers with 30-70% GC content, ensuring an extremely high specificity detection rate. Using 2 µL of 293T cDNA as the template, 1000 pairs of primers for 200 bp amplimers (GC% content 30%-70%) were designed with Primer5 software, and 27 of these amplimers were amplified randomly.
3.High resolution: Can accurately distinguish differences in template concentration of 2-fold
Figure 3. Hieff UNICON™ Universal Blue premix can accurately distinguish differences in template concentration of 2-fold. The template used was a 2-fold gradient dilution of 2 µL of 293T cDNA stock solution, amplifying the GAPDH gene.
4.Good reproducibility of duplicate wells: 90 duplicate wells
Figure 4. Hieff UNICON™ Universal Blue premix exhibits excellent reproducibility, with the amplification curves of 90 duplicate wells showing high coincidence, and a standard deviation of Ct values of less than 0.2. Using 1 µL of 293T cDNA as the template, the GAPDH gene was amplified.
5.Good stability: The premix remains unaffected in amplification performance even after being frozen and thawed 50 times
|
Freeze-thaw cycles |
0 time |
10 times |
30 times |
50 times |
|
Average Ct |
23.11 |
23.17 |
23.18 |
23.25 |
|
▲Ct |
- |
0.06 |
0.07 |
0.14 |
Figure 5. Hieff UNICON™ Universal Blue premix shows no change in amplification performance after being frozen and thawed 50 times. Using 1 µL of 293T cDNA as the template, the C2orf68 gene was amplified with premix that had undergone different numbers of freeze-thaw cycles.
Customer Feedback
Sample source: Mouse ovarian cancer cell line ID8, HM1
Sample source: Mouse epididymal adipose RNA
Sample source: Arabidopsis thaliana leaf RNA
Literature with high impact factor partially published using YEASEN qPCR Mix
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Ordering Information
|
Product Positioning |
Product Name |
Cat# |
|
High Sensitivity Universal Quantitative Premix |
Hieff UNICON™ Universal Blue qPCR SYBR Green Master Mix |
11184ES |
|
High Specificity High Fluorescence Quantitative Premix |
Hieff UNICON™ Advanced qPCR SYBR Master Mix |
11185ES |