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

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-Tn5^{E54K, 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.

SUBMITTER: Soroczynski J

PROVIDER: S-EPMC11257509 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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

Soroczynski Jan J Anderson Lauren J LJ Yeung Joanna L JL Rendleman Justin M JM Oren Deena A DA Konishi Hide A HA Risca Viviana I VI

bioRxiv : the preprint server for biology 20240713

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-Tn5<sup>E54K, L372P</sup> protein per liter of <i>E ...[more]

PMID: 39026714

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