Lipofectamine® 2000 产品资料

Lipofectamine® 2000 Reagent
Cat. nos. 11668-030 Size: 0.3 mL Store at 4°C （do not freeze）
11668-027 0.75 mL
11668-019 1.5 mL
11668-500 15 mL
Pub. Part No. 11668.2k.pps Pub. No. MAN0000995 Rev. Date 20 July 2012
Description
• Lipofectamine® 2000 Transfection Reagent is a proprietary formulation
for transfecting nucleic acids （DNA, RNA, ａnd mRNA） into a wide
range of eukaryotic cells.
• Nucleic acid-Lipofectamine® 2000 complexes can be added directly to
cells in culture medium, in the presence or absence of serum/antibiotic.
• It is not necessary to remove complexes or change/add medium after
transfection.
Important Guidelines for Transfection
• Use Opti-MEM® I Reduced Serum Medium （Cat. no. 31985-062） to
dilute Lipofectamine® 2000 Transfection Reagent ａnd nucleic acids.
• The amount of Lipofectamine® 2000 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® 2000 Reagent to determine an
optimum amount.
• Prepare high-quality plasmid DNA at 0.5–5 μg/μL in deionized water
or TE buffer. A GFP （green fluorescent protein） plasmid can be used to
determine transfection efficiency.
• For additional information, ａnd protocols on transfecting suspension
cells refer to the manual at www.lifetechnologies.com/transfection.
5
Day 2–4 Day 1 Day 0
Cells
Lipofectamine® 2000 Reagent
DNA
Seed cells so they are
70–90% confl uent at the
time of transfection.
Dilute 4 amounts of
Lipofectamine® 2000
Reagent in Opti-MEM®
Medium.
Dilute DNA in
Opti-MEM® Medium.
Add diluted DNA to
each tube of diluted
Lipofectamine® 2000
Reagent （1:1 ratio）.
Incubate.
Add DNA-reagent
complex to cells.
Visualize/analyze
transfected cells.
Plasmid Transfection
Transfect cells according to the following table. The transfection is designed
for 1 DNA amount combined with 4 different amounts of Lipofectamine®
2000. For each lipid reagent amount, the prepared mix is enough to
have triplicates （96-well）, duplicates （24-well）, ａnd single well （6-well）
transfections, ａnd account for pipetting variations.
Component 96-well 24-well 6-well
Adherent cells 1–4 x 104 0.5–2 x 105 0.25–1 x 106
Opti-MEM® Medium 25 μL x 4 50 μL x 4 150 μL x 4
Lipofectamine® 2000 Reagent 1, 1.5, 2, 2.5 μL 2, 3, 4, 5 μL 6, 9, 12, 15 μL
Opti-MEM® Medium 125 μL 250 μL 700 μL
DNA （0.5–5 μg/μL） 2.5 μg 5 μg 14 μg
Diluted DNA 25 μL 50 μL 150 μL
Diluted Lipofectamine® 2000
Reagent 25 μL 50 μL 150 μL
Incubate for 5 minutes at room temperature
DNA-reagent complex/well 10 μL 50 μL 250 μL
Incubate cells for 1–3 days at 37°C
Amount 96-well 24-well 6-well
DNA/well 100 ng 500 ng 2500 ng
Lipofectamine® 2000 Reagent/
well 0.2–0.5 μL 1–2.5 μL 5–12.5 μL
The following table shows the amounts of DNA ａnd Lipofectamine®
2000 Reagent per well used in each transfection reaction. For additional
information on scaling your transfection reaction, see page 4.
For support visit www.lifetechnologies.com/support
www.lifetechnologies.com
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please contact Life Technologies at www.lifetechnologies.com/support.
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