GenElute™ HP Endotoxin-Free Plasmid Maxiprep Kit
SIGMA/NA0410 - sufficient for 25 preparations
Synonym: Gen Elute; GenElute Endotoxin-Free Plasmid Kit
Product Type: Chemical
Assessment of transfection efficiency using plasmid pCop-Green-C.
Plasmid pCop-Green-C is a mammalian expression vector that encodes the copepod green fluorescent protein. HuH-7 cells were seeded onto glass cover slips and then grown to 60-70% confluency. The cells were then transfected with 3 μg of plasmid DNA using ESCORT™ II transfection reagent. Plasmid DNA was isolated from a GenElute™ HP Endotoxin-free Maxiprep. At 72 hours post transfection, the cells were observed under a fluorescence microscope. A) is a picture of the cells following transfection under bright field exposure. B) is the same area of cells under fluorescent exposure, which reveals a high transfection efficiency.
* pCop-Green-C is licensed from Evrogen, JSC.
Plasmid pCop-Green-C is a mammalian expression vector that encodes the copepod green fluorescent protein. HuH-7 cells were seeded onto glass cover slips and then grown to 60-70% confluency. The cells were then transfected with 3 μg of plasmid DNA using ESCORT™ II transfection reagent. Plasmid DNA was isolated from a GenElute™ HP Endotoxin-free Maxiprep. At 72 hours post transfection, the cells were observed under a fluorescence microscope. A) is a picture of the cells following transfection under bright field exposure. B) is the same area of cells under fluorescent exposure, which reveals a high transfection efficiency.
* pCop-Green-C is licensed from Evrogen, JSC.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
| greener alternative category | Aligned  , |
| greener alternative product characteristics | Designing Safer Chemicals Learn more about the Principles of Green Chemistry . |
| Quality Level | 200 |
| storage temp. | 15-25°C |
| sustainability | Greener Alternative Product |
| technique(s) | DNA extraction: suitable |
| usage | sufficient for 25 preparations |
| Application: | GenElute™ HP Endotoxin-Free Plasmid Maxiprep Kit has been used in the isolation of plasmid from swine liver and from competent Escherichia coli cells. |
| Biochem/physiol Actions: | An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is clarified by filtration followed by the addition of a binding solution that has been optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured on silica, while endotoxins are prevented from adsorbing to the membrane. Two wash steps remove contaminants. Finally, the bound DNA is eluted in endotoxin-free water. The recovered plasmid DNA is predominately in its supercoiled form. Genomic DNA and RNA are below detectable levels by ethidium bromide stained agarose gel electrophoresis. |
| Features and Benefits: | • Up to 1.2 mg of high-copy plasmid DNA with endotoxin levels of ≤0.1 EU/μg • Only 35 minutes from pelleted cells to purified plasmid • Convenient vacuum format |
| General description: | Click here to view the kit protocol |
| General description: | The GenElute HP Endotoxin-Free Plasmid Maxiprep Kit offers a simple and rapid method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. The kit uses a vacuum format with a filter column for the rapid clearing of the bacterial lysate and a silica column for capturing plasmid DNA. A proprietary solution binds plasmid DNA to the binding column while preventing endotoxins from adsorbing. The technology allows for the user to consistently achieve levels of endotoxin that are less than 0.1 endotoxin units per μg of plasmid DNA. High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells. |
| Legal Information: | GenElute is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | For additional information, please see www.sigma-aldrich.com/gen. |
| Symbol |     GHS02,GHS05,GHS07,GHS08 |
| Signal word | Danger |
| Hazard statements | H226 - H290 - H302 + H332 - H314 - H334 - H336 - H412 |
| Precautionary statements | P210 - P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 |
| Hazard Codes | C |
| Risk Statements | 20/21/22-32-34-52/53 |
| Safety Statements | 26-36/37/39-45-61 |
| RIDADR | UN 2924 8(3) / PGIII |
| Flash Point(F) | 77.0 °F - closed cup |
| Flash Point(C) | 25 °C - closed cup |
| Supplemental Hazard Statements | EUH032 |
| Storage Temp. | 15-25°C |
| UNSPSC | 41105501 |
| Components | Red Vac Cap 5; Column Preparation Solution 2; Wash Solution 1; Neutralization Solution EF; Wash Solution 2; E-Toxate™ Water, endotoxin, free; Lysis Solution; Resuspension Solution, Add RNase A before use. After addition of RNAse, store at 2-8ºC. ); Binding Solution; RNase A solution |