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STAR Protocols
Volume 3 (1), 2022, 101103

Cloning of DNA oligo pools for in vitro expression

Anna Uzonyi1,2, Ronit Nir1, Schraga Schwartz1,3

Department of Molecular Genetics, Weizmann Institute of Science, 7610001 Rehovot, Israel.

Abstract

Oligo library pools are powerful tools for systematic investigation of genetic and transcriptomic machinery such as promoter function and gene regulation, non-coding RNAs, or RNA modifications. Here, we provide a detailed protocol for cloning DNA oligo pools made up of tens of thousands of different constructs, aiming to preserve the complexity of the pools. This system would be suitable for expression in cell lines and can be followed up by next-generation sequencing analysis.

Keywords: Genetics, Molecular Biology, Systems biology.

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