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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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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