Lipofectamine LTX and Plus Protocol v2.0

Protocol Pub. No. MAN0007822 Rev.1.
Lipofectamine® LTX & PLUS™ Reagent
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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
Selection Lipofectamine® Reagents Scaling Up or Down Transfections

Guide Go online to view related products.
Limited Product Warranty and Disclaimer Details
Product ∤∤Lipofectamine ®
LTX and PLUS™ Reagent are proprietary
formulations for transfecting nucleic acids into a wide range
Description of eukaryotic cells.
∤∤DNA-Lipofectamine LTX and PLUS™ complexes must be

®
made in serum-free medium such as Opti-MEM Reduced®
Serum Medium and can be added directly to cells in culture

medium, in the presence or absence of serum/antibiotic.
Important ∤∤Itmedium
is not necessary to remove complexes or change/add
after transfection.
Guidelines
∤∤The amount of Lipofectamine LTX Reagent required for
®
successful transfection varies depending on the cell type and

passage number. Start any new transfection by testing the
recommended four concentrations of Lipofectamine® LTX
Reagent to determine an optimum amount.
Visit our product page for additional

Online information and protocols. For support,
Resources visit www.lifetechnologies.com/support.
For Research Use Only. Not for use in diagnostic procedures.

Lipofectamine® LTX & PLUS™ Reagent Protocol 2013
Lipofectamine® LTX DNA Transfection Reagents Protocol

Transfect cells according to the following table. Use the indicated volume of DNA and PLUS™ Reagent with each of the four volumes of Lipofectamine® LTX. Each
reaction mix is sufficient for triplicate (96-well), duplicate (24-well), and single well (6-well) transfections, and accounts for pipetting variations. For even less toxicity,
reduce the amount of DNA-lipid complex to the cells, or reduce the amount of DNA used to make complexes.
Timeline Steps Procedure Details

Component 96-well 24-well 6-well
Seed cells to be 70‑90%
Day 0
1 confluent at transfection Adherent cells 1–4 × 104 0.5–2 × 105 0.25–1 × 106
Dilute Lipofectamine® LTX Opti-MEM® Medium 25 μL × 4 50 μL × 4 150 μL × 4

2 Reagent in Opti-MEM®
Lipofectamine® LTX
Medium 1, 1.5, 2, 2.5 μL 2, 3, 4, 5 μL 6, 9, 12, 15 μL
Reagent
Opti-MEM® Medium 125 μL 250 μL 700 μL
Dilute DNA in Opti-MEM ®
Medium, then add PLUS™ DNA (0.5–5 μg/μL) 2.5 μg 5 μg 14 μg

3 Reagent
PLUS™ Reagent 2.5 μL 5 μL 14 μL
Diluted DNA (with PLUS™
Add diluted DNA to diluted 25 μL 50 μL 150 μL
Reagent) Total
Lipofectamine® LTX Reagent
(1:1 ratio) Diluted Lipofectamine® LTX
4 25 μL 50 μL 150 μL
Day 1
Reagent
Incubate Incubate for 5 minutes at room temperature.
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
-2- For support, visit www.lifetechnologies.com/support.

Lipofectamine LTX and Plus Protocol v2.0

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Protocol Pub. No. MAN0007822 Rev.1.

Lipofectamine® LTX & PLUS™ Reagent

Selection Lipofectamine® Reagents Scaling Up or Down Transfections

∤∤DNA-Lipofectamine LTX and PLUS™ complexes must be

made in serum-free medium such as Opti-MEM Reduced®

Serum Medium and can be added directly to cells in culture

successful transfection varies depending on the cell type and

Visit our product page for additional

For Research Use Only. Not for use in diagnostic procedures.

Lipofectamine® LTX DNA Transfection Reagents Protocol

Timeline Steps Procedure Details

Dilute Lipofectamine® LTX Opti-MEM® Medium 25 μL × 4 50 μL × 4 150 μL × 4

Medium, then add PLUS™ DNA (0.5–5 μg/μL) 2.5 μg 5 μg 14 μg

Incubate Incubate for 5 minutes at room temperature.

-2- For support, visit www.lifetechnologies.com/support.

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