Lipofectamine® 2000 Transfection Reagent

（Invitrogen™）

采用Lipofectamine® 2000试剂，您可以实现：

• 在各种细胞系中具有的转染效率和高重组蛋白表达水平 （表1）

• 出众的siRNA和质粒DNA共转染性能

• 在血清存在时亦具有可靠的转染效率 – 转染后无需更换培养基

• 适用于高通量应用的可靠性能

• 建立稳定细胞系的*选择


适用于基因表达和基因沉默的高性能转染试剂

Lipofectamine® 2000试剂适合所有常见细胞系及许多难以转染的细胞系，可用于含有或不含血清的培养基。在基因沉默应用中，Lipofectamine® 2000试剂的高效转染提供了很高的基因敲除水平，足以获得令人信服的结果。Lipofectamine® 2000试剂可实现siRNA和质粒DNA的高效转染，它是共转染的*试剂。Lipofectamine® 2000试剂使用方便 – 只需将其与核酸混合，加入细胞培养物即可。


适合高通量操作

Lipofectamine® 2000试剂使用简单、快速且具有很高的转染效率，适用于瞬时蛋白表达或高通量RNAi转染。在上述应用中，可轻松建立自动化或机器人系统转染条件。如需了解有关Lipofectamine® 2000的实验方案、文献引证及其它产品信息，可登录细胞系数据库。


仅供研究使用。不得用于人或动物的治疗或诊断。

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

Limited Product Warranty

Life Technologies Corporation and/or its affi liate（s） warrant their products as set forth in the Life Technologies’ General Terms and

Conditions of Sale found on Life Technologies’ website at www.lifetechnologies.com/terms and conditions. If you have any questions,

please contact Life Technologies at www.lifetechnologies.com/support.

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*部分 invitrogen 原装产品

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