Getting Started
You’re about to encapsulate a fluid or cytosol! This is how you prepare lipids to be used for liposome encapsulation.
Materials and Equipment
Name | Product | Manufacturer | Part # | Price | Link |
Buffers | |||||
Glucose | D-(+)-Glucose, 99% | Thermo Scientific | A16828-36 | $41.65 | [link] |
Sucrose | Sucrose, 99% | Thermo Scientific | A15583-36 | $41.65 | [link] |
Lipids | |||||
POPC | 16:0-18:1 PC 25 mg/mL | Avanti Lipids | 850457C-500mg | $435.00 | [link] |
Liss-Rhod-PE | 18:0 Liss Rhod PE 1 mg/mL | Avanti Lipids | 810179P-1mg | $273.47 | [link] |
Mineral Oil | Mineral oil, mixed weight | Thermo Scientific | AC415080010 | $53.40 | [link] |
Glass Syringe 250 uL | Hamilton | 14-815-238 | $150.15 | [link] | |
PURE | |||||
PURE | PURExpress | NEB | E6800S | $295.00 | [link] |
RNase Inhibitor | RNase Inhibitor, Murine | NEB | M0314S | $81.00 | [link] |
DNA | pT7-plamGFP PURE | Nucleus v0.1.0-001 DNA | Well E3 | [link] |
Prerequisite Protocols
Protocol
Clean glass syringes.
Pour a small amount of 95% ethanol into a small glass container (10 mL beaker works well).
Assemble the glass syringe and prime it by drawing ethanol into the glass syringe, then empty into a waste bottle.
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Add 3.25mL mineral oil to a glass test tube or beaker.
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Add the following to the mineral oil using the glass syringe, rinsing the syringe with ethanol between each step:
250 uL 25 mg/mL POPC
10 uL 1 mg/mL Liss Rhod PE
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Place lipid beaker in a 55°C dry bath in a fume hood. Leave uncovered for 3 hr to allow chloroform to evaporate.
Let sample cool to room temperature in the fume hood and store in a glass vial until further use.
The lipid/oil mixture can be stored for a week or more at room temperature. Vortex for 1 min before use.
Resources and References
Other Protocols
- The Nucleus protocol simplifies this previous work: Build a Cell Liposome Protocol
Papers
Resources
- Avanti Lipids is a supplier of lipids and offers tips for handling and storage of lipids
Credits
Developers
Testers