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Lipofectamine LTX and Plus Protocol v2.0
Lipofectamine LTX and Plus Protocol v2.0
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Catalog Number Size: LTX/PLUS™
Protocol Outline
Package ∤∤A12621 0.1 mL/40 μL A. Plate cells so they will be 70–90% confluent at the time of
Contents ∤∤15338-030 0.3 mL/0.25 mL transfection.
∤∤15338-100
*PLUS™ Reagent
1.0 mL/0.85 mL (Cat. no. 11514-015) B. Prepare plasmid DNA-lipid complexes.
∤∤15338-500* 15 mL/- available separately. C. Add DNA-lipid complexes to cells.
Lipofectamine® LTX DNA Transfection Reagent Protocol
Storage
Conditions ∤∤Store at 4°C (do not freeze). See page 2 to view a typical plasmid transfection procedure.
∤∤Plasmid DNA (0.5–5 μg/μL stock) Transfection Amounts
Required
∤∤Opti-MEM ®
Reduced Serum Medium
Materials
∤∤Eppendorf tubes Component 96-well 24-well 6-well
DNA per well 100 ng 500 ng 2500 ng
Preparation: 10 minutes
PLUS™ Reagent per well 0.1 μL 0.5 μL 2.5 μL
Timing Incubation: 5 minutes
Final Incubation: 1-3 days Lipofectamine LTX Reagent per well
®
0.2–0.5 μL 1–2.5 μL 5–12.5 μL
Important ∤∤Itmedium
is not necessary to remove complexes or change/add
after transfection.
Guidelines
∤∤The amount of Lipofectamine LTX Reagent required for
®
1 confluent at transfection Adherent cells 1–4 × 104 0.5–2 × 105 0.25–1 × 106
Reagent
5
5 Component 96-well 24-well 6-well
DNA-lipid complex per well 10 μL 50 μL 250 μL
Add DNA-lipid complex
Final DNA used per well 100 ng 500 ng 2500 ng
to cells
Final Lipofectamine® LTX
6 0.2–0.5 μL 1.0–2.5 μL 5.0–12.5 μL
Reagent used per well
Visualize/analyze
Incubate cells for 1–3 days at 37°C. Then, analyze transfected cells.
Day 2–4
transfected cells
7