Published February 4, 2025 | Version v1

Supplementary Data for Chapter "Utilizing Golden Gate Assembly to Streamline CRISPR-Cas/NgTET-Based Phage Mutagenesis"

  • 1. ROR icon Max Planck Institute for Terrestrial Microbiology

Description

This repository complements the book chapter “Utilizing Golden Gate Assembly to Streamline CRISPR-Cas/NgTET-Based Phage Mutagenesis” by Nadiia Pozhydaieva and Katharina Höfer in “Methods in Molecular Biology” (in publishing). It contains the plasmids used for Golden Gate cloning as outlined in the approach. Golden Gate assembly is used to facilitate the exchange of donor DNA sequences (DNA sequences, that are introduced to phage genome upon mutagenesis), to streamline the mutagenesis process and rapidly customize it for various targets. Within this system, the donor DNA is integrated downstream of the gene for NgTET dioxygenase. The latter is crucial for the efficient engineering of phages with extensive cytosine DNA modifications.

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