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bioRxiv · 10.1101/2024.07.11.602973

OpenTn5: Open-Source Resource for Robust and Scalable Tn5 Transposase Purification and Characterization

Soroczynski, J.·Anderson, L. J.·Yeung, J. L.·Rendleman, J. M.·Oren, D. A.·Konishi, H. A.·Risca, V. I.
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Abstract

Tagmentation combines DNA fragmentation and sequencing adapter addition by leveraging the transposition activity of the bacterial cut-and-paste Tn5 transposase, to enable efficient sequencing library preparation. Here we present an open-source protocol for the generation of multi-purpose hyperactive Tn5 transposase, including its benchmarking in CUT&Tag, bulk and single-cell ATAC-seq. The OpenTn5 protocol yields multi-milligram quantities of pG-Tn5E54K, L372P protein per liter of E. coli culture, sufficient for thousands of tagmentation reactions and the enzyme retains activity in storage for more than a year.

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Soroczynski, J., Anderson, L. J., Yeung, J. L., Rendleman, J. M., Oren, D. A., Konishi, H. A., Risca, V. I.. 2024-07-13. OpenTn5: Open-Source Resource for Robust and Scalable Tn5 Transposase Purification and Characterization. https://doi.org/10.1101/2024.07.11.602973

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