• Add transfection complex to the cells in a
dropwise manner.
• Gently shake or swirl the wells or fl asks to
ensure even distribution over the entire plate.
Incubate cells for 18 – 72 hours before
measuring protein expression.
• Place diluent in a sterile tube.
• Add plasmid DNA. Pipet gently to mix.
• Add X-tremeGENE HP DNA Transfection
Reagent to the diluted DNA.
Incubate for 15 min at +15°C to +25°C.
For life science research only. Not for use in diagnostic procedures.
X-TREMEGENE is a trademark of Roche.
Transfection procedureCell preparation for transfection
Plate cells approx. 24 hours before transfection
making sure cells are at optimal concentration
(70 – 90 % confluency).
Steps prior to transfection
Allow X-tremeGENE HP DNA Transfection Reagent,
DNA and diluent (Opti:MEM I Reduced
Serum Medium or serum-free medium) to warm
to +15°C to +25°C, and vortex gently.
A detailed protocol is available in the online instructions for use at www.instructions.roche.com
For cell-type specific protocols, visit www.x-tremegene.roche.com. View the many successfully
used cell-type-specific transfection conditions in our customer feedback database.
Roche Diagnostics GmbH
Sandhofer Straße 116
68305 Mannheim
Germany
© 2011 Roche Diagnostics.
All rights reserved.
06516084001
0911
X-tremeGENE HP DNA Transfection Reagent
Quick Protocol
For products with Catalog Numbers:
06 365 752 001 | 06 366 244 001 | 06 366 236 001 | 06 366 546 001
Volumes of X-tremeGENE HP DNA Transfection Reagent and amounts of DNA for various ratios
Ratio
Transfection Reagent : DNA
Minimal transfection
complex volume
Volume for a whole
96-well plate
Serum-free medium to a final volume of
100 µl 500 µl
1 : 1
X-tremeGENE HP Transfection Reagent
1 µl 5 µl
DNA
1 µg 5 µg
2 : 1
X-tremeGENE HP Transfection Reagent
2 µl 10 µl
DNA
1 µg 5 µg
3 : 1
X-tremeGENE HP Transfection Reagent
3 µl 15 µl
DNA
1 µg 5 µg
A C
B