Gentaur plasmids

The protocols for working with Gentaur plasmids involve tasks such as plasmid DNA extraction, transformation into host cells, and analysis. Here’s a basic advice of Gentaur for procedures associated with plasmid work:

Plasmid DNA Extraction:

Materials Needed:

• Bacterial culture containing plasmids
• Plasmid DNA extraction kit
• Centrifuge
• Microcentrifuge tubes
• Isopropanol and ethanol
• DNA storage buffer

Procedure:

1. Bacterial Culture:
   • Grow a bacterial culture containing plasmids.
2. Harvest Cells:
   • Centrifuge the culture to collect bacterial cells.
3. Resuspend Cells:
   • Resuspend the bacterial pellet in a lysis buffer.
4. Lysis:
   • Add an alkaline lysis solution to lyse the cells and denature proteins.
5. Neutralization:
   • Neutralize the lysate to stabilize plasmid DNA.
6. Centrifugation:
   • Centrifuge to separate cell debris from the cleared lysate.
7. DNA Precipitation:
   • Precipitate DNA by adding isopropanol.
8. Washing:
   • Wash the DNA pellet with ethanol.
9. DNA Resuspension:
   • Resuspend the purified plasmid DNA in a storage buffer.

Plasmid Transformation:

Materials Needed:

• Competent cells (bacterial cells made permeable for DNA uptake)
• Plasmid DNA
• Transformation buffer
• Heat shock apparatus
• Recovery medium (LB broth or SOC medium)

Procedure:

1. Thaw Competent Cells:
   • Thaw competent cells on ice.
2. Add DNA:
   • Add plasmid DNA to competent cells.
3. Incubation:
   • Incubate the cells and DNA on ice.
4. Heat Shock:
   • Subject the cells to a brief heat shock.
5. Cooling:
   • Cool the cells on ice again.
6. Recovery:
   • Add recovery medium and incubate the cells at a suitable temperature.
7. Plating:
   • Plate cells on selective agar plates.
8. Incubation:
   • Incubate the plates overnight for colony growth.

Plasmid Analysis:

Materials Needed:

• Agarose gel
• Gel electrophoresis apparatus
• DNA ladder
• Ethidium bromide or other DNA stain
• UV transilluminator

Procedure:

1. Prepare Agarose Gel:
   • Cast an agarose gel with appropriate concentration.
2. Load Samples:
   • Mix plasmid DNA samples with loading dye and load onto the gel.
3. Run Electrophoresis:
   • Run the gel at a suitable voltage to separate DNA fragments.
4. Staining:
   • Stain the gel with ethidium bromide or another DNA stain.
5. Visualization:
   • Visualize DNA bands under a UV transilluminator.

Gentaur offers a wide range of plasmids that match your research needs:

https://maxanim.com/genetics/plasmid/