X-tremeGENE™ Transfection Reagents General Recommendations

X-tremeGENE™ HP and X-tremeGENE™ 9 Transfection Reagent Comparison

The ability to successfully transfect a cell varies significantly depending on the cell type, protocol, and the composition of the transfection reagent. X-tremeGENE™ transfection reagents are designed to efficiently transfect common and difficult-to-transfect cells with a variety of molecules, including DNA, small RNAs, and CRISPR/Cas9 components.

 

  X-tremeGENE™ HP DNA Transfection Reagent   X-tremeGENE™ 9 DNA Transfection Reagent  
Transfection of DNA
Efficiency with standard cell lines (e.g., COS-7, HEK-293, HeLa, NIH 3T3, and CHO-K1)
Efficiency with "difficult-to-transfect" cells (e.g., HT 29, HCT 116, and K-562 cells)
Transfection of primary cells NR
Gentleness of the reagent
Ease of use (minimal optimization)
Cytotoxicity after transfection Very low low

NR: Not Recommended 

 

Focus Application X-tremeGENE™ HP DNA Transfection Reagent X-tremeGENE™ 9 DNA Transfection Reagent X-tremeGENE™ siRNA Transfection Reagent
Functional studies of cellular processes
Quantitative protein expression
Gene knockdown studies using siRNA

X-tremeGENE™ HP Reagent efficiently transfects human primary fibroblasts

Figure 1: X-tremeGENE HP Reagent efficiently transfects human primary fibroblasts.

Primary fibroblasts were isolated from human foreskin and transfected with 4 μl X-tremeGENE HP Reagent and 1 μg of GFP-encoding plasmid DNA in a 6-well cell culture plate. GFP expression was visualized 48 hours after transfection. The left panel shows GFP fluorescence, and the right panel shows the corresponding brightfield image.

 

X-tremeGENE™ HP and 9 Reagents outperform the competitor reagent in human mesenchymal stem cells

Figure 2: X-tremeGENE HP and 9 Reagents outperform the competitor reagent in human mesenchymal stem cells.

Human mesenchymal stem cells were isolated from bone marrow aspirate of four different donors. Cells were transfected using X-tremeGENE HP Reagent (XHP), X-tremeGENE 9 Reagent (X9), and a competitor reagent (LTX), using the indicated ratios (μl reagent : μg GFP-encoding plasmid DNA. Ratio 2:1 was not tested using X-tremeGENE 9 Reagent). Untransfected cells served as control (neg). FACS analysis was performed 48 hours after transfection.

  X-tremeGENE™ HP DNA Transfection Reagent outperforms competitor reagents in difficult-to-transfect cell types.

Figure 3: X-tremeGENE HP DNA Transfection Reagent outperforms competitor reagents in difficult-to-transfect cell types.

Cells were transfected using X-tremeGENE HP Reagent (XHP), and competitor reagents (LTX and L2K), at the indicated ratios (μl reagent : μg GFP-encoding plasmid DNA). Transfection efficiency was visualized using GFP fluorescence microscopy.

Cell-Type Specific Product Recommendations

Established Cell Lines X-tremeGENE™ HP DNA Transfection Reagent X-tremeGENE™ 9 DNA Transfection Reagent
CHO-K1
COS-7
NIH-3T3
HeLa
HEK-293
Hep-G2
HT1080
HT29 NR
PC-3
K-562 NR
U-2 OS NR
MCF-7
RAW NR
HCT 116
Jurkat NR
PC-12
Primary Cells    
Human primary fibroblasts NR
MEF NR
Stem Cells    
Human mesenchymal stem cells NR
Mouse embryonic stem cells (E14TG2a)
Insect Cell Lines    
High Five NR
SF9 NR

NR: Not Recommended

Note: The above data was produced using either a GFP-encoding pcDNA3.1 plasmid or a luciferase-encoding pCI plasmid, both with the cytomegalovirus (CMV) promoter. These recommendations are guidelines based on experimental findings. The optimal reagent:DNA ratio must be determined empirically.

Application Table

Focus Applications Recommended Product Advantages
  • Cancer research
    Transfection of common cancer research
    cell lines for functional cellular analysis, including
    gene regulation and protein interaction studies
    and pathway analysis.

  • Target evaluation
    Transfection of common cell lines for target
    evaluation.
  • Low cytotoxicity
  • Minimal reagent-induced off-target effects
  • Simple method
  • Broad applicability across cell types
  • High transfection efficiency at low cell density
  • Active in up to 100% serum
  • Difficult-to-transfect and primary cells
    Transfection of primary cells and "hard-to-transfect"
    cell lines for functional cellular analysis.

  • Protein expression
    Transfection for analytical or quantitative
    protein expression.
  • High levels of protein expression
  • Simple method
  • Rapid protocol
  • High transfection efficiency
  • Active in serum-containing media
  • Synthetic; free of human- or animal-derived components
  • Gene knockdown using siRNA transfection
  • Low cytotoxicity
  • High knockdown efficiency
  • Broad applicability across cell lines
  • Active in serum-containing media

 

X-tremeGENE is a trademark of Roche. For life science research only. Not for use in diagnostic procedures.