For life science research only.

Not for use in diagnostic procedures.

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X-tremeGENE HP DNA Transfection Reagent

For transient and stable transfection of eukaryotic cells

Cat. No. 06 365 752 001Cat. No. 06 366 244 001Cat. No. 06 366 236 001Cat. No. 06 366 546 001Trial-pack0.4 ml1 ml5 × 1 ml

Version 05

Content version: May 2011

Store at –15 to –25°C

1.What this Product Does

Number of Tests

Using the standard procedure, 1 ml of X-tremeGENE HP DNA Trans-fection Reagent can be used to perform up to 10,000 transfections in 96-well plates.

Formulation

X-tremeGENE HP DNA Transfection Reagent is a proprietary blend of lipids and other components supplied in 80% ethanol, filtered through 0.2 m pore size membrane, and packaged in glass vials. It does not contain any ingredients of human or animal origin.

Storage and Stability

Store X-tremeGENE HP DNA Transfection Reagent at –15 to –25°C, with the lid tightly closed. The reagent is stable until the expiration date printed on the label when stored under these conditions.

LX-tremeGENE HP DNA Transfection Reagent remains fully func-tional even after repeated opening of the vial (at least five times over a two-month period), as long as the vial is tightly recapped and stored at -15 to -25°C.

LNote that the shipping temperature of this product is different

from the storage temperature. These different temperatures will not affect product performance or product stability.Special Handling

NAfter removing the amount required, tightly close the vial with the

lid immediately after use.

NAlways bring the vial to +15 to +25°C and mix X-tremeGENE HP

DNA Transfection Reagent prior to removing the amount required vortexing for one second.

NDo not aliquot X-tremeGENE HP DNA Transfection Reagent; store

in the original glass vials.

NMinimize the contact of undiluted X-tremeGENE HP DNA Trans-fection Reagent with plastic surfaces.

NFor use, the minimum amount of X-tremeGENE HP DNA Transfec-tion Reagent: DNA complex is 100 µl. Complex formation at lower volumes can significantly decrease transfection efficiency.

NDo not use tubes or microplates made of polystyrene for

X-tremeGENE HP Transfection Reagent : DNA complex prepara-tion. When not able to avoid polystyrene materials, make certain to pipet the transfection reagent directly into the serum-free medium (e.g., Opti-Mem).

NDo not use siliconized pipette tips or tubes.

Additional Equipment and Reagents Required

Additional reagents and equipment required to perform transfection assays using X-tremeGENE HP DNA Transfection Reagent include: Standard Laboratory Equipment.

Standard cell culture equipment (e.g., biohazard hoods, incuba-tors)

Standard pipettes and micropipettes Vortex mixer

For Plasmid Preparation

Purified plasmid stock (0.1 – 2.0 µg/µl) in sterile TE (10 mM Tris, 1 mM EDTA, pH 8.0) buffer or sterile water

Genopure Plasmid Midi Kit* or Genopure Plasmid Maxi Kit* to prepare plasmid

For Verification of Vector Function Assay appropriately for transfected gene

G-418 Solution* or Hygromycin B* (optional for stable transfec-tion experiments)

For Transfection-Complex Formation

Opti-MEM I Reduced Serum Medium or serum-free medium Sterile polypropylene tubes or round-bottom 96-well plates Growing Cells

Select subconfluent cultures in log phase for preparation of cell cultures

Quantify cell number to reproducibly plate the same number of cellsApplication

X-tremeGENE HP DNA Transfection Reagent is a high performance transfection reagent, free of animal-derived components. Benefits of X-tremeGENE HP DNA Transfection Reagent include:

Designed to transfect a broad range of eukaryotic cells, including insect cells, many cell lines not transfected well by other reagents, and hard-to-transfect cell lines (e.g., HT-1080, K-562, HepG2).

Can be successfully used in a variety of applications, such as gene expression analysis and protein production using transiently trans-fected cells, generation of stable cell lines, expression of shRNA for gene knockdown studies, drug discovery programs, and target evaluation. Samples and detailed transfection protocols are avail-able at www.51wendang.com.

Produces minimal cytotoxicity or changes in morphology when ade-quate numbers of cells are transfected, eliminating the requirement to change media after adding the transfection complex. Suitable for transient and stable transfection.

Functions very well in the presence or absence of serum.