Transfection reagent: Roche [shopping book]

Nowadays, a number of new products have emerged in the transfection market, and some old products are more stringent. When it comes to Roche's transfection reagents, everyone is sure to think of Fugene 6/HD.

Even if Fugene 6/HD is not in hand, Roche is not afraid because it combines 20 years of transfection experience with X-tremeGENE 9 and X-tremeGENE HP transfection reagents . X-tremeGENE 9 reagents provide high performance for a variety of commonly used cell lines, while X-tremeGENE HP reagents are more suitable for difficult-to-transfect cells, such as primary cells and insect cells. One of the two reagents is similar to Fugene, which is mild to cells.

X-tremeGENE 9 & X-tremeGENE HP

Both reagents combine low cytotoxicity with high transfection efficiency, allowing more cells to survive and bring more physiologically relevant results. Moreover, the transfection step is also quite simple: simply dilute the transfection reagent with serum-free medium, incubate with the plasmid, and then add the cells dropwise. Researchers need very little optimization, of course, you still have to optimize the ratio of transfection reagent to DNA before transfection. The recommended ratio is 3:1, but for a particular study, a ratio of 2:1 to 7:1 may increase transfection efficiency.

In addition, X-tremeGENE HP reagent is also filtered through a 0.2 μm pore size membrane, free of animal-derived components, and active in serum. It forms a complex with nucleic acids that promotes efficient DNA transfer with minimal physical and morphological changes. It is ideal for cell analysis and protein expression studies.

How to choose these two reagents, please see the table below. If you transfect HeLa, HEK-293 and other common cell lines, both can do the job. For those difficult to get cells, it is best to choose X-tremeGENE HP reagent. If you talk about the mildness of the reagents, I am afraid that X-tremeGENE 9 reagents will be better.

The price of X-tremeGENE 9 reagent is 1764 yuan / 0.4 ml, 3545 yuan / 1 ml, and the price of X-tremeGENE HP reagent is 2098 yuan / 0.4 ml, 4851 yuan / 1 ml. (price is for reference only)

Use X-tremeGENE 9/HD tips

1. X-tremeGENE 9 DNA transfection reagent should be stored at +2 to +8 °C, and X-tremeGENE HP DNA transfection reagent should be stored at -15 to -25 °C.
2. Place the glass bottle containing X-tremeGENE DNA Transfection Reagent at +15 to +25 °C and vortex for one second before use.
3. Do not dispense X-tremeGENE DNA transfection reagent; store the remaining transfection reagent in the original glass bottle.

4. Try to avoid contact of the undiluted X-tremeGENE DNA transfection reagent with the plastic surface.
5. When used, the minimum volume of X-tremeGENE DNA transfection reagent and DNA complex is 100μl. Reducing the volume of the complex will significantly reduce transfection efficiency.
6. Avoid the preparation of X-tremeGENE DNA transfection reagents and DNA complexes using polystyrene tubes or microplates. If polystyrene materials cannot be avoided, the transfection reagent should be added directly to serum-free medium or Opti-MEM I low serum medium.
7. Do not use (poly) silicone treated tips or tubes.

8. Ensure that cells are still experiencing exponential growth during the transfection period.
9. At the time of transfection, appropriate controls should be designed, including: (1) untransfected cells; (2) cells to which only transfection reagents are added; and (3) cells to which only DNA is added.

10. The optimal ratio of transfection reagent to DNA, and the optimal total volume of the complex, should be adjusted based on cell line, cell density, cell growth status at the time of detection, and gene expression.

X-tremeGENE siRNA Transfection Reagent

X-tremeGENE siRNA Transfection Reagent is a liposome-based, optimized transfection reagent. Although the name says siRNA, it can also be used for co-transfection of siRNA and expression plasmids. It forms a complex with a small interfering RNA (siRNA) or siRNA/plasmid DNA mixture and efficiently transfects into animal cells, triggering corresponding gene silencing. Relatively small intracellular toxicity ensures that the observed cellular effects are only associated with transfected or co-transfected siRNA. Transfection can be performed with or without serum, and no transfusion operation is required after transfection.

Protocol: X-tremeGENE siRNA transfection reagent and siRNA were diluted in serum-free Opti-MEM, mixed and incubated for 15-20 minutes at room temperature. Add cells and monitor the interference effects.
Price: 2816 yuan / 1 ml, 11981 yuan / 5 × 1 ml (recommended dosage of 2.5 μl per hole in a 24-well plate) (price is for reference only)

Tips for using X-tremeGENE siRNA

1. For the initial optimization, the following amounts of X-tremeGENE siRNA transfection reagent (μl) and siRNA (μg) were used: 10:2, 2.5:0.5, 1:0.2.
2. According to Roche's experience, 2.5 μl of X-tremeGENE siRNA transfection reagent is used in each well of a 24-well plate to achieve efficient knockdown of most cell types.
3. If possible, exclude antibacterial agents during the initial experiment. Once efficient conditions are established, these components can be added back when monitoring the transfection results. X-tremeGENE Reagent: The siRNA complex must be prepared in serum-free medium, even if the cells are transfected in the presence of serum.

Request more information on X-tremeGENE transfection reagents!

Cell-specific transfection step

Roche offers a number of cell-specific specific protocols as a guide to your use of transfection reagents. Of course, these are for reference only. Roche also has a transfection database that provides links to relevant citations and some customer feedback. You can search for the relevant transfection steps based on the cell name, and it is convenient to use the drop-down menu. For example, when searching for 293 cells transfected with X-tremeGENE 9 transfection reagent, the following results appear on the web page.

293 cells have been successfully transfected with 7 clients using X-tremeGENE 9 transfection reagent and plasmid DNA.
Transfection type: The best results were obtained in the following ratios (reagent μl:DNA μg): 3:1; 2:1
Note: These percentages from customer feedback are for guidance only and have not been verified by Roche. The optimal transfection conditions may vary and should be determined empirically.

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