GeneRulor Transposase

1. Product Overview

GeneRulor Transposase is a highly active transposase engineered by molecular directed evolution, originating from the Tn5 transposase in Escherichia coli. Its transposition efficiency is significantly improved compared with the native transposase through protein engineering optimization. The enzyme can specifically recognize DNA fragments containing Mosaic End (ME) sequences with high specificity and assemble with them to form a ready-to-use transposome complex with pre-embedded adapters, which can be directly used for transposition reactions and simplifies the experimental procedure.

Its mechanism of action is that the transposase can recognize the inverted repeats equences at the ends of the transposon and form a synaptic complex. In the presence of magnesium ions (Mg²⁺), it completes the cleavage of double-stranded DNA and adapter ligation simultaneously. This characteristic enables it to efficiently and randomly insert the carried DNA fragments into the target sequence, and exhibits low sequence preference for DNA samples with different GC contents, ensuring the wide applicability and stability of the transposition reaction. Therefore, this enzyme is particularly suitable for molecular biology applications such as high-throughput sequencing library construction.

2. Product Features

(1) Ultra-high activity: The transposition efficiency is 5-10 times higher than that of thenative enzyme after protein engineering optimization.

(2) Broad substrate compatibility: Extremely low sequence preference for DNA samples with different GC contents.

(3) High specificity: Precisely recognizes ME sequences to minimize non-specificreactions.

(4) Batch-to-batch consistency: A strict quality control system ensuresstable performance between batches.

3. Applications

3.1 Constructionof bacterial transposon mutant libraries

(1) Transposome formation: Incubate GeneRulor Transposase with DNA fragments containing ME sequences in Assemble Buffer.

(2) Electroporation: Electroporate the reaction product into recipient bacterial strains and obtain transposon insertion mutants through antibiotic screening.

(3) Mutant library construction: Collect individual clones to form a mutant pool, which can be directly used for functional screening or subsequent NGS analysis.

Figure 1. Schematic diagram of transposon design

3.2 Next-Generation Sequencing (NGS) Library Construction

GeneRulor Transposase is a highly efficient tool for constructing DNA libraries for next-generation sequencing. It is characterized by utilizing the special transposition mechanism of the GeneRulor Transposase dimer to complete two steps (cleavage of double-stranded DNA and adapter ligation) simultaneously. This enzyme features rapid reaction and low sample requirement, and can fragment DNA and add specific adapter sequences to both ends in a single reaction. This simplified "one-step" tagging method greatly optimizes the experimental procedure, which not only saves operation time but also reduces experimental costs, thus being widely used in the field of DNA sequencing library construction.

Figure 2. Schematic diagram of the workflow fornext-generation sequencing library construction

References

[1] Li N, Jin K, Bai Y, Fu H, LiuL, Liu B. Tn5 Transposase Applied in Genomics Research. Int J Mol Sci. 2020 Nov6;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.