Title

Design of customizable long linear DNA substrates with controlled end modifications for single-molecule studies

RIS ID

140996

Publication Details

Mueller, S., Spenkelink, L., van Oijen, A. & Lewis, J. (2020). Design of customizable long linear DNA substrates with controlled end modifications for single-molecule studies. Analytical Biochemistry, 592

Abstract

2019 Many strategies have been developed to manipulate DNA molecules and investigate protein-DNA interactions with single-molecule resolution. Often, these require long DNA molecules with a length of several 10s of kb that are chemically modified at specific regions. This need has traditionally been met by commercially available DNA from bacteriophage λ. However, λ DNA does not allow for the generation of highly customizable substrates in a straightforward manner, an important factor when developing assays to study complex biochemical reactions. Here we present a generalizable method for the design and production of very long chemically modified DNA substrates derived from a single plasmid. We show the versatility of this design by demonstrating its application in studying DNA replication in vitro. We anticipate this strategy will be broadly useful in producing a range of long chemically modified DNA molecules required for a diverse range of single-molecule approaches.

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Link to publisher version (DOI)

http://dx.doi.org/10.1016/j.ab.2019.113541