Cat.No.：D15011A
Price: ￥
Specifications：
This product adopts the SYBR Green I hybrid fluorescence method and is a highly integrated 2× qPCR premix solution. The kit contains all the reaction components such as hot-start Taq DNA polymerase, optimized buffer, dNTPs, Mg²⁺, and SYBR Green I dye. The core principle lies in that the SYBR Green I dye specifically binds to double-stranded DNA and releases fluorescence, and the fluorescence signal intensity is strictly proportional to the amplification amount of the target gene. This product is easy to operate, has high amplification efficiency, and can precisely meet the real-time quantitative analysis requirements of various target genes.
High sensitivity and high efficiency: The optimized buffer system ensures low Ct values and high amplification efficiency (90% – 110%), significantly enhancing the detection sensitivity.
Strong specificity guarantee: Relying on the thermal start technology, it significantly reduces primer dimerization and non-specific amplification interference, ensuring accurate results.
Wide dynamic linear range: Supports a wide linear range from single copy to 10⁹ copies, meeting the quantitative needs of various samples.
Melting curve analysis: Supports melting curve analysis after amplification, easily verifying the specificity of the product and eliminating false positive interference.
Full instrument compatibility: Strong versatility, low consumable cost, seamlessly compatible with all mainstream brands of fluorescence quantitative PCR instruments on the market.
| Cat. No. | Component | Specification | Quantity |
| B20011A | Dye-based qPCR Master Mix (2×) | 1 mL | 1 |
Store at -20℃ and transport at ≤0℃
Gene mRNA relative expression quantification, strain and microbiota quantification, gene copy number variation analysis, and routine fluorescence quantitative experiments in research.
The amplification curve shows abnormalities. What could be the reason for this?
The common amplification abnormalities can be classified into the following situations: 1. The curve is not smooth or has a jagged appearance: This may be due to weak fluorescence signal, degradation of the dye in the reagent, or improper addition of the reference dye (such as ROX). It is recommended to ensure that the dyes in the Mix have not degraded, to replace the qPCR-specific consumables with better fluorescence signal collection capabilities, or to correct the reference dye. 2. The curve breaks, declines, or suddenly drops sharply: If the template concentration is high and the baseline endpoint value is greater than the Ct value, it will cause the curve to break or decline. You can try to reduce the baseline endpoint (Ct value - 4) and re-analyze the data; if the curve in a particular well drops sharply, it is usually caused by bubbles remaining in the reaction tube, which burst after the temperature rises. After adding the sample, carefully check and centrifuge to remove the bubbles. 3. The curve is "S"-shaped: This is usually caused by excessive template concentration in the reaction system, poor nucleic acid quality, or the presence of inhibitors, resulting in deviation in amplification efficiency (either too high or too low). It is recommended to remove the template with the highest concentration and re-analyze.
There was no Ct value obtained during the amplification. What is the reason for this? Why?
Possible causes include incorrect settings in the fluorescence signal acquisition step (for example, the two-step method should collect data during the annealing/extension stage, while the three-step method should collect data at 72°C during the extension stage), degradation of primers or probes, insufficient template quantity or severe degradation of the template. It is recommended to check the instrument settings, conduct electrophoresis to detect the integrity of the primers, and start testing unknown concentration samples from the highest concentration.
