Literature DB >> 32728517

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

Hyun-Min Kim1, Sisi Tian1, Shouyue Wang1.   

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. © King Abdulaziz City for Science and Technology 2020.

Entities:  

Keywords:  CRISPR-Cas9; Genome editing; Microinjection; Plasmid miniprep

Year:  2020        PMID: 32728517      PMCID: PMC7376781          DOI: 10.1007/s13205-020-02346-7

Source DB:  PubMed          Journal:  3 Biotech        ISSN: 2190-5738            Impact factor:   2.406


  16 in total

1.  CRISPR-Cas9-Guided Genome Engineering in Caenorhabditis elegans.

Authors:  Hyun-Min Kim; Monica P Colaiácovo
Journal:  Curr Protoc Mol Biol       Date:  2019-12

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Journal:  J Biol Chem       Date:  1988-09-25       Impact factor: 5.157

6.  Purification of plasmid and BAC transgenic DNA constructs.

Authors:  Chengyu Liu; Yubin Du; Wen Xie; Changyun Gui
Journal:  Methods Mol Biol       Date:  2013

7.  Lipopolysaccharide is a frequent contaminant of plasmid DNA preparations and can be toxic to primary human cells in the presence of adenovirus.

Authors:  M Cotten; A Baker; M Saltik; E Wagner; M Buschle
Journal:  Gene Ther       Date:  1994-07       Impact factor: 5.250

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Authors:  S Brenner
Journal:  Genetics       Date:  1974-05       Impact factor: 4.562

9.  Caenorhabditis elegans innate immune response triggered by Salmonella enterica requires intact LPS and is mediated by a MAPK signaling pathway.

Authors:  Alejandro Aballay; Eliana Drenkard; Layla R Hilbun; Frederick M Ausubel
Journal:  Curr Biol       Date:  2003-01-08       Impact factor: 10.834

10.  Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes.

Authors:  Adam D Norris; Hyun-Min Kim; Mónica P Colaiácovo; John A Calarco
Journal:  Genetics       Date:  2015-07-30       Impact factor: 4.562

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