GeneRulor Tn5-Plus Transposase

1. Product Overview

With its unique "cut-and-paste" mechanism, Tn5 transposase can complete DNA fragmentation and adapter ligation in one step, shortening the traditional library construction process from several hours to about 10 minutes and reducing the required initial DNA amount to the ng level. Therefore, it has become a core tool for technologies including Whole Genome Sequencing (WGS), ATAC-seq, CUT&Tag, transcriptome analysis and single-cell sequencing. Nevertheless, the inherent sequence bias of Tn5 transposase has become a key bottleneck restricting the quality and uniformity of sequencing data.

To address this common industry challenge, GeneRulor has developed a new-generation transposase, GeneRulor Tn5-Plus Transposase. Based on the three-dimensional structure of the Tn5-DNA complex, this product has undergone rational design and multiple rounds of directed evolution, with precise modification to its target DNA-binding domain. While maintaining and further enhancing core catalytic activity, GeneRulor Tn5-Plus Transposase significantly reduces insertion sequence bias, thus achieving more uniform and authentic genome coverage and providing a more reliable data foundation for various high-throughput sequencing applications.

2. Product Features

2.1 Precise Design

Systematic optimization is focused on the target DNA-binding domain, while the structural integrity of the ME sequence recognition region and catalytic active center is fully considered. This not only significantly improves the functional performance of the enzyme but also ensures the stability and reliability of its core catalytic mechanism.

2.2 Significantly Reduced Insertion Sequence Bias

Quantitative evaluation of sequence insertion sites by Shannon entropy shows that Tn5-Plus exhibits a significant reduction in sequence bias. As a key indicator for measuring sequence randomness, a higher Shannon entropy value indicates that the distribution of insertion sites is closer to the ideal random state. Experiments have demonstrated that compared with wild-type Tn5, Tn5-Plus has a more uniform insertion distribution in various genomic backgrounds, effectively reducing coverage bias caused by sequence preference. This improvement fundamentally enhances the unbiasedness and reliability of sequencing data, providing a more authentic and consistent genomic mapping foundation for subsequent bioinformatics analysis.

2.3 More Balanced Whole-Genome Coverage

When processing AT-rich regions in the genome, wild-type Tn5 transposase often suffers from low insertion efficiency, leading to insufficient coverage of these regions, which become blind spots and sources of bias in deep sequencing. Tn5-Plus significantly enhances the recognition and insertion capabilities in these high-AT regions, thereby greatly improving the fragmentation efficiency of such regions. This improvement makes the distribution of insertion events more balanced across the whole genome, effectively reducing data bias caused by sequence preference and providing more comprehensive and authentic genome coverage information for downstream analysis.

2.4 Higher Fragmentation Activity

Under the same reaction conditions, Tn5-Plus has a higher fragmentation efficiency than wild-type Tn5. The distribution of its product fragments is highly concentrated, mainly in the ideal library construction range of 200–500 bp, which is conducive to improving the effective data output of sequencing libraries and providing a library construction tool with better performance for high-throughput sequencing.

3. Application

Owing to its comprehensively optimized performance, GeneRulor Tn5-Plus Transposase is not only fully compatible with all application scenarios of wild-type Tn5, but also able to provide a more reliable data foundation for various NGS applications with lower sequence bias, higher fragmentation efficiency and more stable enzymatic activity. It is especially suitable for key scenarios with stringent requirements for data uniformity and integrity:

(1) Whole Genome Sequencing (WGS): Its more uniform DNA fragmentation capability can effectively reduce coverage bias caused by genomic factors such as GC content, yielding whole-genome data with more even coverage.

(2) ATAC-seq: In the research field of chromatin open regions, its low sequence bias enables more authentic and comprehensive capture of genome-wide accessible sites, effectively reducing background noise and improving detection sensitivity.

(3) Single-cell sequencing: Faced with extremely limited starting samples, its higher enzymatic activity stability and cleavage efficiency can ensure efficient and uniform library construction even under single-cell or ultra-low input conditions, preserving biological information to the maximum extent.

(4) Transposon library construction: As a core tool enzyme, its optimized enzymatic properties enable efficient and random fragmentation and adapter ligation of target DNA (e.g., plasmids, genomes, PCR products, etc.), facilitating the construction of mutant or sequencing libraries with more complete coverage.

(5) Transcriptome sequencing and other high-throughput library construction: In applications such as RNA-seq, its low bias helps achieve a more uniform distribution of cDNA fragments and improve the accuracy of gene expression quantification, making it particularly suitable for the detection of complex transcripts or low-abundance transcripts.

References

[1] Li N, Jin K, Bai Y, Fu H, Liu L, Liu B. Tn5 Transposase Applied in Genomics Research. Int J Mol Sci. 2020 Nov 6;21(21):8329. doi: 10.3390/ijms21218329.

[2] Zhang H, Lu T, Liu S, Yang J, Sun G, Cheng T, Xu J, Chen F, Yen K. Comprehensive understanding of Tn5 insertion preference improves transcription regulatory element identification. NAR Genom Bioinform. 2021 Oct 27;3(4):lqab094. doi: 10.1093/nargab/lqab094.