Lipid Preparation

<aside> <img src="/icons/light-bulb_gray.svg" alt="/icons/light-bulb_gray.svg" width="40px" /> Getting Started

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You’re about to encapsulate a fluid or cytosol! This is how you prepare lipids to be used for liposome encapsulation.

<aside> <img src="/icons/wrench_gray.svg" alt="/icons/wrench_gray.svg" width="40px" /> Materials and Equipment

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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	16:0 Liss Rhod PE 1 mg/mL	Avanti Lipids	810158C-1mg	$281.67	[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]

<aside> <img src="/icons/merge_gray.svg" alt="/icons/merge_gray.svg" width="40px" /> Prerequisite Protocols

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<aside> <img src="/icons/iterate_gray.svg" alt="/icons/iterate_gray.svg" width="40px" /> Protocol

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[ ] 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.
Tips
[ ] Add 3.25mL mineral oil to a glass test tube or beaker.
Tips
[ ] 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
Tips
[ ] 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.

<aside> <img src="/icons/book_gray.svg" alt="/icons/book_gray.svg" width="40px" /> Resources and References

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<aside> <img src="/icons/document_gray.svg" alt="/icons/document_gray.svg" width="40px" /> Other Protocols

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The Nucleus protocol simplifies this previous work: Build a Cell Liposome Protocol

<aside> <img src="/icons/book-closed_gray.svg" alt="/icons/book-closed_gray.svg" width="40px" /> Papers

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<aside> <img src="/icons/info-alternate_gray.svg" alt="/icons/info-alternate_gray.svg" width="40px" /> Resources

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Avanti Lipids is a supplier of lipids and offers tips for handling and storage of lipids

<aside> <img src="/icons/megaphone_gray.svg" alt="/icons/megaphone_gray.svg" width="40px" /> Credits

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<aside> <img src="/icons/search_gray.svg" alt="/icons/search_gray.svg" width="40px" /> Developers

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<aside> <img src="/icons/verified_gray.svg" alt="/icons/verified_gray.svg" width="40px" /> Testers

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