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Saturation Mutagenesis Genome Engineering of Infective ΦX174 Bacteriophage via Unamplified Oligo Pools and Golden Gate Assembly.

ACS synthetic biology - 17 Jan 2020

Faber Matthew S, Van Leuven James T, Ederer Martina M, Sapozhnikov Yesol, Wilson Zoë L, Wichman Holly A, Whitehead Timothy A, Miller Craig R

Abstract excerpt

Here we present a novel protocol for the construction of saturation single-site-and massive multisite-mutant libraries of a bacteriophage. We segmented the ΦX174 genome into 14 nontoxic and nonreplicative fragments compatible with Golden Gate assembly. We next used nicking mutagenesis with oligonucleotides prepared from unamplified oligo pools with individual segments as templates to prepare near-comprehensive...

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Identifiers

PMID: 31825605
DOI: 10.1021/acssynbio.9b00411
PMCID: PMC7055157

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PubMed metadata and Literature Corpus relationships. Corpus snapshot updated 7 Sept 2026. Manifest publication-graph-cohort-2026-09-07.1.

Source receipt: pubmed, retrieved 7 Sept 2026.

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