NGS Library Amplification New Choice | 2× Fidelis Master Mix High-Fidelity Hot-Start Pre-Mixed Solution

Author: Senior Product Manager of UA Views：6

2× Fidelis Master Mix is specifically designed for NGS library construction, integrating ultra-high fidelity, rapid hot-start amplification, long-fragment extension capability, and compatibility with complex templates. It effectively reduces amplification error rates, shortens experimental cycles, and enhances system stability, providing efficient and reliable domestic enzymatic tools for genomics research.

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Recent Advances

UniA 2× Fidelis Master Mix Provides a High-Fidelity Amplification Solution for NGS Library Construction.

Brief Description: 2× Fidelis Master Mix is specifically designed for NGS library construction, integrating ultra-high fidelity, rapid hot-start amplification, long-fragment extension capability, and compatibility with complex templates. It effectively reduces amplification error rates, shortens experimental cycles, and enhances system stability, providing an efficient and reliable domestic enzymatic tool for genomics research.

I. Next-generation sequencing technology imposes stringent requirements on the performance of library amplification enzymes.

With the rapid development of Next-Generation Sequencing (NGS) technology, research in fields such as genomics, transcriptomics, and epigenetics has set unprecedented high standards for the quality of sequencing data. In the NGS library construction workflow, PCR amplification of target fragments is one of the key steps determining final data quality. This step not only requires amplifying trace amounts of DNA template to the load capacity required by the sequencer, but also must ensure that the amplification product accurately reflects the sequence information of the original template. Therefore, erroneous bases (mutations) introduced during PCR amplification will directly lead to false-positive variants in sequencing data, interfering with downstream bioinformatics analysis. To address this technical bottleneck, UniA has launched a PCR high-fidelity amplification solution specifically designed for NGS. Its core product, 2× Fidelis Master Mix, with its outstanding enzymatic properties, provides reliable technical support for high-precision library construction.

II. Ultra-high fidelity is the core cornerstone for ensuring NGS data accuracy.

During library amplification, the fidelity of DNA polymerase is the primary indicator for evaluating its performance. Poster data show that the fidelity of 2× Fidelis Master Mix is approximately 90 times that of conventional Taq DNA polymerase. This significant improvement originates from directed engineering of its core enzyme, which possesses extremely strong 3'→5' exonuclease activity (i.e., proofreading function), enabling it to effectively identify and excise mismatched bases. More critically, this product has been empirically validated through sequencing of 288 clones, and the results show that almost no mismatched bases are incorporated into its amplification products. In academic research, this means that for complex genomes or low-frequency mutation detection (such as tumor liquid biopsy), this master mix can minimize the noise introduced by PCR amplification to the greatest extent, ensuring the sensitivity and specificity of variant detection, thereby laying a solid enzymatic foundation for the production of high-quality sequencing data.

III. The rapid and efficient hot-start system significantly optimizes amplification kinetics.

In addition to fidelity, amplification efficiency and time cost are also key concerns for researchers. 2× Fidelis Master Mix employs high-fidelity hot-start technology. By blocking enzyme activity at room temperature through antibody or chemical modification, this system effectively inhibits non-specific amplification and primer dimer formation, greatly improving reaction specificity. In terms of amplification kinetics, this system demonstrates the advantage of being "fast enough": its 98°C denaturation time requires only 20 seconds, and extension speed can reach 30-60 seconds/kb. In a conventional PCR program, the entire amplification process can be completed in about 1 hour. This rapid amplification feature not only shortens the experimental cycle and improves the efficiency of high-throughput sample processing, but the hot-start mechanism also ensures stability when preparing reaction systems at room temperature, reducing the impact of operational errors on experimental results.

IV. Excellent long-fragment amplification capability meets the needs of complex library construction.

NGS library construction sometimes needs to cover larger genomic regions or handle long-fragment templates. 2× Fidelis Master Mix possesses "long enough" amplification capability. For long-fragment templates such as 1.6 kb, 2.5 kb, and 3.4 kb, this master mix can amplify bright bands, demonstrating its powerful long-fragment extension capability. For ultra-long templates greater than 10 kb, this solution also provides a dedicated protocol, further broadening the product's application boundaries. This long-fragment amplification capability benefits from its highly processive DNA polymerase core, which can complete the synthesis of long-chain DNA within a relatively short time without easily dissociating, thereby meeting experimental scenarios with strict requirements for long-fragment amplification, such as complex structural variation research or full-length gene cloning.

V. Excellent system stability and compatibility ensure experimental reproducibility.

When dealing with high-GC-content templates or difficult templates with complex secondary structures, PCR amplification often faces challenges. 2× Fidelis Master Mix demonstrates the characteristic of being "stable enough." Its formulation contains an optimized buffer system and provides a supporting DMSO addition protocol for high-GC or difficult templates. As a commonly used PCR enhancer, DMSO can disrupt DNA secondary structures and lower the melting temperature, thereby promoting primer-template binding. This master mix also exhibits calm stability when preparing reactions at room temperature, meaning researchers do not need to operate on ice, reducing operational difficulty and improving experimental reproducibility. This compatibility with difficult templates and convenience of operation make the product broadly applicable to library construction from various complex samples.

VI. Comprehensive specifications and service system facilitate one-stop advancement of research work.

2× Fidelis Master Mix is available in two specifications, 1.25 mL and 5*1.25 mL, to meet the needs of experiments with different throughputs. As an independent domestic brand under UniV, UniA relies on the service philosophy of "Drug Discovery, One-Stop Solution" to provide researchers with a complete solution from reagents to technical support. Users can obtain detailed product information by scanning the QR code on the poster, or contact UniV sales to apply for a trial and request a quote. As a high-fidelity enzyme dedicated to NGS, this product strictly follows the ISO quality management system, ensuring batch-to-batch stability and providing a cost-effective domestic alternative for genomics research in China, helping researchers in the life sciences to efficiently and accurately complete their scientific exploration.

This article is reviewed and published by the technical expert team of UA

Disclaimer: This article partially utilizes artificial intelligence assistance in its creation. If any content involves copyright or intellectual property issues, please let us know and we promise to verify and remove it as soon as possible.

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