Cloning of DNA oligo pools for in vitro expression

Anna Uzonyi; Ronit Nir; Schraga Schwartz

doi:10.1016/j.xpro.2021.101103

Cloning of DNA oligo pools for in vitro expression

STAR Protoc. 2022 Jan 17;3(1):101103. doi: 10.1016/j.xpro.2021.101103. eCollection 2022 Mar 18.

Authors

Anna Uzonyi¹, Ronit Nir¹, Schraga Schwartz¹

Affiliation

¹ 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. For complete details on the use and execution of this profile, please refer to Uzonyi et al. (2021).

Keywords: Genetics; Molecular Biology; Systems biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cloning, Molecular
DNA* / genetics
Gene Library
High-Throughput Nucleotide Sequencing*

Substances

DNA