Biolistic-Mediated Gene Transfer: A Technique to Deliver Gene of Interest in Target Cells via a Biolistic Gene Gun
Published: April 30, 2023
Abstract
Source: Taylor, T., et al. Biolistic Transformation of a Fluorescent Tagged Gene into the Opportunistic Fungal Pathogen Cryptococcus neoformans. J. Vis. Exp. (2015).
In this video, we demonstrate the protocol of biolistic gene gun-mediated gene delivery in cultured cells. This method provides a simple method of stable genetic transformation using a minimal quantity of DNA.
Protocol
1. DNA Preparation Prepare orange macrocarrier biolistic discs by submerging them in 100% ethanol using forceps. Place discs into a large petri dish containing drierite to dry (make sure the drierite does not touch the discs). Once dry, press the macrocarrier discs into the silver disc holders (previously wiped down with 100% ethanol). Vortex gold microcarrier beads suspended in 100% ethanol and aliquot 12 µl into a 1.5 ml microcentrifuge tube, one tube per transforma…
Divulgaciones
The authors have nothing to disclose.
Materials
| 0.6 μm gold beads | Bio-Rad | 165-2262 | http://www.bio-rad.com |
| Spermadine-free base | Sigma- Aldrich | S0266 | https://www.sigmaaldrich.com |
| 1350 psi Rupture Discs | Bio-Rad | 165-2330 | http://www.bio-rad.com |
| Stopping Screens | Bio-Rad | 165-2336 | http://www.bio-rad.com |
| Macrocarriers discs | Bio-Rad | 165-2335 | http://www.bio-rad.com |
| PDS-1000/He System | Bio-Rad | 165-2257 | http://www.bio-rad.com |
| Microfuge 18 Centrifuge | Beckman Coulter | F241.5P | www.beckmancoulter.com |
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Biolistic-Mediated Gene Transfer: A Technique to Deliver Gene of Interest in Target Cells via a Biolistic Gene Gun. J. Vis. Exp. (Pending Publication), e20996, doi: (2023).