Методы Именно локализованного передачи ячеек или ДНК в начале постимплантационных эмбрионов мыши
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Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| Forceps | Dumostar | T5390 | |
| Dissecting stereomicroscope | Zeiss | Stemi 2000-C | |
| Stereomicroscope system with fluorescence | Nikon | AZ100 | |
| Inverted microscope with a digital camera | Olympus | Olympus BX61 | |
| Inverted confocal microscope | Leica Microsystems | Leica TCS SP8 | |
| Low melting point agarose | Life Technologies | 16520-050 | |
| Pasteur pipettes | Fisher Scientific | 11397863 | |
| 30mm Petri dishes | Fisher Scientific | 121V | |
| 4-well plates | Thermo scientific | 179820 | |
| M2 medium | Sigma-Aldrich | M7167 | |
| Phosphate Buffered Saline (PBS) | Life Technologies | 10010015 | |
| Paraformaldehyde (PFA) | Sigma-Aldrich | P6148 | |
| Pipettes | NICHIRYO | Nichipet | |
| tips | Greiner Bio One | 685280 | |
| Cell culture incubator | SANYO | MCO-17AIC | |
| Roller culture apparatus | BTC Engineering | ||
| Syringe filters 0.45µm, sterile | Sigma-Aldrich | 10462100 | |
| Glasgow Minimum Essential Medium (GMEM) | Sigma-Aldrich | G5154 | |
| non-essential amino acids (NEAA) | Life Technologies | 11140050 | |
| L-glutamine | Fisher Scientific | SH30549.01 | |
| Sodium pyruvate solution | Fisher Scientific | SH30239.01 | |
| Penicillin and Streptomycin 10.000UI/ml | Lonza | DE17-602E | |
| Gas Cartridge for Portable Meker Burner | COLEMAN | COLEMAN 250 | |
| Thin Wall Borosilicate Capillary Glass with Fillament, OD 1.0 mm, ID 0.78 mm | Harvard Apparatus | 640798 | |
| Aspirator tube assemblies for calibrated microcapillary pipettes | Sigma-Aldrich | A5177-5EA | |
| Flaming/Brown micropipette puller | Sutter Instrument Company | P-97 | |
| Microforge | De Fonbrune | BS030301 | |
| Pneumatic pico pump | World Precision Instruments | PV830 | |
| Microloader tips | Eppendorf | 5242956.003 | |
| ECM 830 square wave pulse generator | BTX | 45-0002 | |
| Green food coloring dye | Sigma-Aldrich | C.I. 42053 | |
| A far-red cell membrane-impermeable nuclear dye | Biotium | 40060-T | |
| pCAG-Cre:GFP | Addgene | #13776 | |
| pCAG-GFP | Addgene | #16664 | |
| Multimeter | Excel | XL830L | |
| Micromanipulators | Leitz | ||
| 0.2mm diameter platinum wire | Agar Scientific | E404-2 | |
| Anti-GFP antibody | Abcam | ab13970 | |
| Goat anti-Chicken IgY, HRP | Santa Cruz | sc-2428 | |
| Liquid DAB+ Substrate Chromogen System | Dako | K3467 | |
| 4',6-diamidino-2-phenylindole (DAPI) | Life Technologies | D21490 | |
| A far-red fluorescence nuclear counterstain | Life Technologies | T3605 |
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Cite This Article
Huang, Y., Wilkie, R., Wilson, V. Methods for Precisely Localized Transfer of Cells or DNA into Early Postimplantation Mouse Embryos. J. Vis. Exp. (106), e53295, doi:10.3791/53295 (2015).