The plasmids used in this study were: pCag-DsRed (Addgene #11151; 0.7 μg/μL) and pCAG_GPHN.FingR-eGFP-CCR5TC (Addgene #46296; 0.8 μg/μL; Gross et al., 2013 (link)). The DNA was purified with a Maxiprep Endofree Kit (Macherey-Nagel, Düren, Germany), and diluted in 1x phosphate-buffered saline (PBS). The DNA solution (with 0.05% Fast green) was injected into the lateral ventricle of E14.5 embryos using pulled glass pipettes. The embryos were electroporated using tweezers-type electrodes (CUY650-5). Five square electric pulses were passed (38 V, 50 ms interval cycle length, 950 ms interval pause).
Maxiprep endofree kit
Manufactured by Macherey-Nagel
Sourced in Germany
The Maxiprep Endofree Kit is a laboratory tool designed for the isolation and purification of high-quality plasmid DNA from bacterial cultures. The kit utilizes a modified alkaline lysis method and provides an efficient way to obtain endotoxin-free plasmid DNA suitable for various downstream applications.
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Lab products found in correlation
2 protocols using maxiprep endofree kit
1
In vivo plasmid electroporation
2
In Utero Electroporation of Plasmids
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Plasmids used in this study were as follows: pCag-DsRed (Addgene, #11151; 2 µg/µL), pCag-GFP (Addgene, #11150; 1 µg/µL), pCALNL-GFP (Addgene, #13770; 1 µg/µL), and pCag-Cre (Addgene, #13775; 4 ng/µL) (62 (link), 63 (link)). Plasmids were purified with a Maxiprep Endofree Kit (Macherey-Nagel) and diluted in 1x PBS. pCag-DsRed was added to the mix of electroporation as a control of efficiency. The DNA solution (with 0.05% Fast-green) was injected into the lateral ventricle of E15.5 embryos using pulled glass pipettes. The embryos were electroporated using tweezer-type electrodes (CUY650-5). Five square electric pulses were passed (40V, 50-ms interval cycle length, 950-ms interval pause). Electroporated animals, expressing DsRed, were selected using a fluorescent flashlight lamp (Nightsea, DFP-1).
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