Microinjection of Expression Vectors

Ac, phiC31 integrase, and FLP expression vectors (Figure 1A) were linearized using NotI, purified using phenol-chloroform extraction, and used as template to synthesize capped mRNAs using the Message mMachine SP6Kit (Life Technologies, Gaithersburg, MD) as described previously (Ishikawa et al. 2013 (link)). The RNAs were purified using an RNeasy MinElute (Qiagen) kit and microinjected into one-cell-stage embryos at the indicated concentration. Microinjection was performed as described previously (Ishikawa et al. 2011 (link)).

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Ishikawa T., Ansai S., Kinoshita M, & Mori K. (2018). A Collection of Transgenic Medaka Strains for Efficient Site-Directed Transgenesis Mediated by phiC31 Integrase. G3: Genes|Genomes|Genetics, 8(8), 2585-2593.

Publication 2018

Message mMachine SP6Kit RNeasy MinElute

Cell Chloroform Embryos Integrase Microinjection Mrnas Phenol Rnas Vectors

Corresponding Organization :

Other organizations : Kyoto University

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Variable analysis

independent variables

Ac
PhiC31 integrase
FLP expression vectors

dependent variables

Not specified

control variables

Concentration of RNAs microinjected into one-cell-stage embryos

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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