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Reliable, reagent-based DNA transfection for eukaryotic cells using calcium phosphate co-precipitation
The Invitrogen™ Calcium Phosphate Transfection Kit provides a convenient and effective method for introducing plasmid DNA into mammalian cell lines using the classic calcium phosphate co-precipitation technique. This time-tested method enables both transient expression and stable transformation of a wide range of eukaryotic cell types.
The kit includes all necessary components, pre-formulated and quality-tested, to generate fine DNA-calcium phosphate precipitates that efficiently adhere to and enter cells via endocytosis. It is ideal for research laboratories working with established adherent cell lines in standard plate or flask formats.
How It Works
DNA is mixed with 2 M CaCl₂ and then added dropwise into a HEPES-buffered phosphate solution, forming fine calcium phosphate–DNA precipitates. These precipitates attach to the cell surface, facilitating DNA uptake by endocytosis. The process is optimized by aeration of the buffer during addition to ensure a fine precipitate, which enhances transfection efficiency.
Key Features
- Reliable and Reproducible: Classic and widely adopted method for stable and transient transfection
- Complete Kit: Includes calcium chloride, HEPES buffer, sterile water, and a control plasmid (pcDNA™3.1/His/lacZ)
- Optimized Formulation: Produces fine, uniform precipitates that promote efficient DNA uptake
- Flexible Use: Suitable for a range of well plate and flask formats
- Manual Protocol: Ideal for small- to mid-scale research applications
Applications
- Transient gene expression in mammalian cells
- Generation of stable transfectants
- Functional gene studies
- Plasmid delivery into established cell lines
- Basic research in molecular and cellular biology
Kit Components & Storage
| Component | Volume |
|---|---|
| Tissue Culture Sterile Water | 2 × 12 mL |
| 2X HEPES Buffered Saline (HBS) | 2 × 12 mL |
| 2 M Calcium Chloride (CaCl₂) | 3 × 1 mL |
| Control Plasmid: pcDNA™3.1/His/lacZ | 20 µg |
Storage Conditions: Store all components at –5 °C to –30 °C
Technical Specifications
| Specification | Details |
|---|---|
| Catalog Number | K278001 |
| Format | Compatible with flasks and 6-, 12-, 24-, 48-, 96-well plates |
| Transfection Technique | Chemical (Calcium Phosphate Co-precipitation) |
| Sample Type | Plasmid DNA |
| Serum Compatible | No |
| Cell Type | Established mammalian cell lines |
| High-throughput Compatible | No |
| Product Type | Transfection Kit |
| Quantity | 75 Reactions |
| Unit Size | Each |
The Invitrogen™ Calcium Phosphate Transfection Kit offers a dependable, economical, and well-established method for DNA transfection in mammalian cells. With all critical reagents included and optimized for performance, it is ideal for academic and research laboratories focused on gene function, expression analysis, and stable cell line generation.
User Guide
Reliable, reagent-based DNA transfection for eukaryotic cells using calcium phosphate co-precipitation
The Invitrogen™ Calcium Phosphate Transfection Kit provides a convenient and effective method for introducing plasmid DNA into mammalian cell lines using the classic calcium phosphate co-precipitation technique. This time-tested method enables both transient expression and stable transformation of a wide range of eukaryotic cell types.
The kit includes all necessary components, pre-formulated and quality-tested, to generate fine DNA-calcium phosphate precipitates that efficiently adhere to and enter cells via endocytosis. It is ideal for research laboratories working with established adherent cell lines in standard plate or flask formats.
How It Works
DNA is mixed with 2 M CaCl₂ and then added dropwise into a HEPES-buffered phosphate solution, forming fine calcium phosphate–DNA precipitates. These precipitates attach to the cell surface, facilitating DNA uptake by endocytosis. The process is optimized by aeration of the buffer during addition to ensure a fine precipitate, which enhances transfection efficiency.
Key Features
- Reliable and Reproducible: Classic and widely adopted method for stable and transient transfection
- Complete Kit: Includes calcium chloride, HEPES buffer, sterile water, and a control plasmid (pcDNA™3.1/His/lacZ)
- Optimized Formulation: Produces fine, uniform precipitates that promote efficient DNA uptake
- Flexible Use: Suitable for a range of well plate and flask formats
- Manual Protocol: Ideal for small- to mid-scale research applications
Applications
- Transient gene expression in mammalian cells
- Generation of stable transfectants
- Functional gene studies
- Plasmid delivery into established cell lines
- Basic research in molecular and cellular biology
Kit Components & Storage
| Component | Volume |
|---|---|
| Tissue Culture Sterile Water | 2 × 12 mL |
| 2X HEPES Buffered Saline (HBS) | 2 × 12 mL |
| 2 M Calcium Chloride (CaCl₂) | 3 × 1 mL |
| Control Plasmid: pcDNA™3.1/His/lacZ | 20 µg |
Storage Conditions: Store all components at –5 °C to –30 °C
Technical Specifications
| Specification | Details |
|---|---|
| Catalog Number | K278001 |
| Format | Compatible with flasks and 6-, 12-, 24-, 48-, 96-well plates |
| Transfection Technique | Chemical (Calcium Phosphate Co-precipitation) |
| Sample Type | Plasmid DNA |
| Serum Compatible | No |
| Cell Type | Established mammalian cell lines |
| High-throughput Compatible | No |
| Product Type | Transfection Kit |
| Quantity | 75 Reactions |
| Unit Size | Each |
The Invitrogen™ Calcium Phosphate Transfection Kit offers a dependable, economical, and well-established method for DNA transfection in mammalian cells. With all critical reagents included and optimized for performance, it is ideal for academic and research laboratories focused on gene function, expression analysis, and stable cell line generation.
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