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[Preprint]. 2024 Jul 13:2024.07.11.602973. [Version 1] doi: 10.1101/2024.07.11.602973

Figure 1: OpenTn5 enables robust, scalable, open-source Tn5 transposase purification for multiple applications.

Figure 1:

A. Diagram of simultaneous DNA fragmentation and addition of DNA sequencing adapters in vitro by Tn5 transposition reaction, referred to as tagmentation. B. Expression and purification of pG-Tn5 can be carried out within 5 days and yields multi-milligram amounts of enzyme, sufficient for thousands of tagmentation reactions. Less than 1.25 μg of pG-Tn5 is required per 50’000 cell input ATAC-seq reaction. C. Schematic of 10His-pG-Tn5(E54K, L372P) expression plasmid, VR124 (Addgene #198468). D. Robust over-expression of soluble 10His-pG-Tn5(E54K, L372P) protein. E. His-tag affinity and size exclusion chromatography (SEC) enable efficient purification of pG-Tn5 homodimer. F. Comparison of pG-Tn5 adapter unloaded stock at 1 mg/mL with commercial Illumina TDE1 enzyme (Cat #20034198). G. Comparison of tagmentation activity of pG-Tn5 (1 mg/mL) and Illumina TDE1 using equal transposome volumes on Lambda genomic DNA (λ gDNA).