Journal: Nature protocols

Article Title: Germline transgenesis in rodents by pronuclear microinjection of Sleeping Beauty transposons.

doi: 10.1038/nprot.2014.008

Figure Lengend Snippet: Figure 2 | Result of in vitro mRNA synthesis and test of the microinjection buffer. mRNA quality can be determined by standard RNase-free agarose electrophoresis and ethidium bromide staining. Samples are run on a 1% (wt/vol) RNase-free nondenaturing agarose gel. Lane M, DNA size marker; lane 1, 1 µl of in vitro–synthesized mRNA; lane 2, 1 µl of in vitro– synthesized mRNA incubated (1 h, 37 °C) in 10 µl of EmbryoMax injection buffer to detect the potential presence of RNase in the buffer (reproduced with permission from ref. 80). Shown is a result without visible signs of mRNA degradation.

Article Snippet: mMessage mMachine T7 kit (Invitrogen/Ambion, cat. no. AM1344) Plasmid DNA preparation kit (Qiagen, cat. no. 27106) Agarose, DNase, RNase none detected (Sigma-Aldrich, cat. no. A4718) Ethidium bromide, 1% (wt/vol) solution in water (Sigma-Aldrich, cat. no. E7637) ! cautIon This is a hazardous chemical.

Techniques: In Vitro, Microinjection, Electrophoresis, Staining, Agarose Gel Electrophoresis, Marker, Synthesized, Incubation, Injection