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An affordable plasmid miniprep suitable for proficient microinjection in Caenorhabditis elegans.

ABSTRACT: A variety of techniques, including CRISPR-Cas9 genome editing, have been developed to produce genetically modified cell lines and animal models. In many cases, the success of the genome-editing techniques is dependent on the quality of the introduced DNA. However, the preparation of high-quality plasmids required for small-scale microinjection has not been explored. Here, we compared various types of plasmid preparation methods for their microinjection proficiency and developed an efficient and affordable plasmid mini preparation method suitable for Caenorhabditis elegans microinjection. By combining the advantages of Triton X-114 and column-based mini preparation (hence, we named it TXC), the new TXC method was affordable, efficient, and equivalent to expensive plasmid midiprep method based on microinjection efficiency. Besides, TXC was compatible with general molecular biology grade reactions and worked proficiently for different types of plasmids.

SUBMITTER: Kim HM

PROVIDER: S-EPMC7376781 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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An affordable plasmid miniprep suitable for proficient microinjection in Caenorhabditis elegans.

Kim Hyun-Min HM Tian Sisi S Wang Shouyue S

3 Biotech 20200722 8

A variety of techniques, including CRISPR-Cas9 genome editing, have been developed to produce genetically modified cell lines and animal models. In many cases, the success of the genome-editing techniques is dependent on the quality of the introduced DNA. However, the preparation of high-quality plasmids required for small-scale microinjection has not been explored. Here, we compared various types of plasmid preparation methods for their microinjection proficiency and developed an efficient and ...[more]

PMID: 32728517

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