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Imunoanalýza
Cas9 Expressing Immortalized Chicken Embryonic Fibroblast (DF1) Cell Line
| Unit |
1×10⁶ cells / 1.0 ml |
|---|---|
| Cell Type |
Stable (KO/Expressing) Cells |
| Tissue Type |
Embryo |
| Organism |
Chicken (Galline) |
| Donor History | |
| Growth Properties |
Adherent ,polygonal |
| Growth Conditions |
PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. Dulbecco's Modified Eagle Medium (DMEM) ,High Glucose (TM500) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255) ,37.0°C ,5% CO₂. |
| Immortalization Method | |
| Full Information |
https://www.abmgood.com/cas9-expressing-immortalized-chicken-embryonic-fibroblast-df1-cell-line.html |
To generate the Cas9 Expressing Immortalized Chicken Embryonic Fibroblast (DF1) Cell Line, Cas9-2A-PuroR was cloned into pT2KCAGGS using Gibson Assembly (NEB). The DF1 cells were then transfected, and selected for using puromycin. The constitutive nuclear expression of Cas9 was validated by immunostaining using an antibody against Cas9, followed by DAPI-counterstaining. This DF1 cell line can be used to greatly streamline the process of experimental designs for future CRISPR/Cas9 knockout experiments in chick embryos.
