lipofectamine2000_man

Lipofectamine® 2000 Reagent

Cat. nos. 11668-030 Size: 0.3 mL 11668-027 0.75 mL 11668-019 1.5 mL 11668-500 15 mLU

Pub. Part No. 11668.2k.pps

Pub. No. MAN0000995

Store at 4°C (do not freeze)

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

se 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 http://doc.guandang.net/transfection.

For research use only. Not for human or animal therapeutic or diagnostic use.

Cells

yaDLipofectamine® 2000 Reagent

DNA

1 yaD

4–2 yaD

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.

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.

Scaling Up or Down Transfections

determine the reagent amount needed for your new plate format.2

Optimum amount needed is determined from the protocol (see pages 2–3).

Co-Transfection of Plasmid DNA and siRNA

Transfect plasmid DNA and siRNA at the same time using Lipofectamine® 2000 Reagent by adding 30 pmol (~0.6 μg) of siRNA per 1 μg of DNA.

mRNA Transfection

mRNA can be transfected in a 24-well plate by using Lipofectamine® 2000 Reagent by adding 0.5–1 μg of mRNA per well.

Limited Product Warranty

Life Technologies Corporation and/or its af liate(s) warrant their products as set forth in the Life Technologies’ General Terms and Conditions of Sale found on Life Technologies’ website at http://doc.guandang.net/terms and conditions. If you have any questions, please contact Life Technologies at http://doc.guandang.net/support.

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