Thermal-initiated dTTP (100 mM solution)

Commodity name： Thermal-initiated dTTP (100 mM solution)
货号： OLV1405

Category:

Hot-start dNTPs

Email:

wangtongtong@cnorilife.com

Phone:

+86-15032236037

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Product Description

As a core technology in the field of in vitro diagnostics, PCR testing has seen significant advancements in recent years, driven by evolving market demands. To ensure timely and accurate test results during large-scale outbreaks and control efforts—such as those related to COVID-19 and African swine fever—IVD companies have increasingly begun developing fully pre-mixed products. Major raw-material suppliers have also promptly updated their formulations at the raw-material level. Nevertheless, the severe issue of non-specific amplification in fully pre-mixed reagents continues to plague researchers, remaining an intractable challenge. Addressing the non-specificity problem in fully pre-mixed reagents has now become one of the major challenges in the field of molecular biology. Currently, the sealing efficiency of hot-start DNA polymerases is only around 90%–95%. Once primers, probes, enzymes, and buffers are all pre-mixed together, any enzyme activity that remains unsealed can lead to substantial non-specific amplification in the reaction system. To overcome this shortcoming of hot-start enzymes and completely eliminate the issue of non-specific amplification, Olive Bio proudly introduces its revolutionary new product: Thermal-Initiated dNTP—“Thermal-initiated dNTP Mix”!
Product Introduction

The Thermal-initiated dNTP Mix is a revolutionary, innovative product developed by Olive Bio Company. It is a specially chemically modified deoxynucleoside triphosphate that, under normal conditions, cannot participate in the formation of phosphodiester bonds. Only after being subjected to thermal activation does the Thermal-initiated dNTP Mix transform into a conventional dNTP and become capable of participating in PCR amplification. This product can significantly reduce nonspecific amplification caused by non-specific annealing or primer dimers.

The thermally initiated dNTP Mix can be stably stored at -20°C for at least one year. It is compatible with a variety of hot-start DNA polymerases and is suitable for various PCR systems, including conventional PCR, rapid thermal-cycling PCR, multiplex PCR, and real-time fluorescent quantitative PCR. When used in combination with a hot-start DNA polymerase, it can minimize non-specific amplification to the greatest extent.

Introducing the Thermal-Initiated dNTP Monomer Kit—perfectly tailored to meet the application needs of downstream manufacturers!

Product Features

1. Good sealing performance at room temperature

To verify the sealing effect of the thermal-initiated dNTP Mix, we chose to conduct the sealing efficacy test in a constant-temperature PCR (LAMP) system, where the temperature throughout the entire PCR process does not exceed 60℃.
- When the all-premixed system was left standing for 3 days, the results showed that, compared to conventional dNTPs, the thermally initiated dNTP Mix exhibited complete closure with no detectable signal (Figure 1-A, blue box—no heating).
- After the thermal-initiated dNTP components were individually preheated at 95℃ for 30 seconds, their amplification performance was comparable to that of conventional dNTPs (Figure 1-B).
Figure 1-A: Schematic illustration of the sealing effect of the Thermal-initiated dNTP Mix under a constant-temperature PCR system.

Figure 1-B: Comparison of amplification efficiency between thermally activated dNTPs and conventional dNTPs in the LAMP system after the thermal-initiated dNTPs have been pre-activated separately.

2. High release efficiency after heat shock

Under temperature-shift PCR conditions, the Thermal-initiated dNTP Mix does not require separate heating and can be fully released during a standard PCR protocol. Despite its rapid release kinetics, the Thermal-initiated dNTP Mix maintains amplification efficiency comparable to that of conventional dNTPs (as shown in Figure 2A). Under isothermal LAMP conditions, when the components of the Thermal-initiated dNTP Mix are heated separately for just 30 seconds, their sealing effect can be quickly reversed, and the amplification efficiency is nearly identical to that of conventional dNTPs (as shown in Figure 2B).

Figure 2: Comparison of amplification results between the Thermal-initiated dNTP Mix and conventional dNTPs under a temperature-gradient PCR system.

3. Effectively reduces the occurrence of non-specific amplification.

To investigate the effect of the thermal-initiated dNTP Mix in reducing nonspecific amplification, we conducted Capillary Electrophoresis Experiment Under the STR super-multiplex system, we compared the Thermal-initiated dNTP Mix with conventional dNTP products. The results showed that the thermal-initiated DNA polymerase, when used in combination with conventional dNTPs, led to substantial nonspecific amplification (indicated by the red box in Figure 4, lower panel). In contrast, the Thermal-initiated dNTP Mix, when used together with the thermal-initiated DNA polymerase, significantly reduced nonspecific amplification (as shown in the red box area in Figure 4).

Figure 3: Comparison of capillary electrophoresis results between the thermally initiated dNTP Mix and the conventional dNTP Mix.