Staphylococcus aureus RN4220 strain is commonly used in studies involving virulence, resistance, metabolism and more.
Staphylococcus aureus (S. aureus) RN4220 Electrocompetent Cells
$705.00
Description
Intact Genomics (ig®) Staphylococcus aureus (S. aureus) RN4220 Electrocompetent Cells are optimized to provide high transformation efficiencies making them ideal for various applications. This strain is commonly used in studies involving virulence, resistance, metabolism and more. Staphylococcus aureus RN4220 is characterized by a mutation in the sau1 hsdR genes, making it restriction deficient and hence an excellent intermediate cloning host.
Specifications
Competent cell type: Electrocompetent
Species: S. aureus
Strain: RN4220
Format: Tubes
Transformation efficiency: ≥1 x 105 cfu/µg DNA
Blue/white screening: No
Shipping condition: Dry ice
Reagents Included
ig® Staphylococcus aureus (S. aureus) RN4220 Electrocompetent Cells
Recovery medium
Storage
ig® Staphylococcus aureus (S. aureus) RN4220 Electrocompetent Cells: -80 ºC
Recovery medium: 4 ºC
Quality Control
Transformation efficiency is tested by using the control DNA supplied with the kit and using the high-efficiency transformation protocol. Transformation efficiency should be equal or greater than 1×105 CFU/µg DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
Follow these guidelines when using ig® S. aureus Electrocompetent Cells:
• Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical
lysis caused by pipetting.
• Thaw competent cells on ice and transform cells immediately following thawing. After adding DNA,
mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.
Biosafety Level: 2
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories. 5th ed. Washington, DC: U.S. Government Printing Office, 2009; see:
www.cdc.gov/biosafety/publications/bmbl5/index.htm.
BSL2 facility is required for purchase and use of this product 1294-40
Additional information
| µl | 5×200µl |
|---|
Transformation Protocol
Use this procedure to transform ig® Staphylococcus aureus ElectroCompetent Cells. Do not use these cells for chemically transformation.
- Place sterile cuvettes and microcentrifuge tubes on ice.
- Remove competent cells from the -80 °C freezer and thaw completely on wet ice (10-15 minutes).
- Aliquot 1 µl (10pg -1 µg) of DNA to the chilled microcentrifuge tubes on ice.
- When the cells are thawed, add 25 μl of cells to each DNA tube on ice and mix gently by tapping 4-5 times. For the control, add 1 µl of (50 ng/µl) DNA to the 25 µl of cells on ice. Mix well by tapping. Do not pipette up and down or vortex to mix, this can harm cells and decrease transformation efficiency.
- Pipette 26 µl of the cell/DNA mixture into a chilled electroporation cuvette without introducing bubbles. Quickly flick the cuvette downward with your wrist to deposit the cells across the bottom of the well and then electroporate.
- Immediately add 500µl of Recovery Medium or any other medium of choice to the cuvette, pipette up and down three times to re-suspend the cells. Transfer the cells and Recovery Medium to an Eppendorf tube.
- Incubate tubes at 37 °C for 1 hours at 200 RPM.
- Dilute the cells as appropriate then spread 20-200 μl cells onto a pre-warmed selective plate. For the control plasmid , you may plate 100 μl of undiluted transformation mix onto a YT plate containing chloramphenicol 12.5µg/µl). Use sterilized spreader or autoclaved ColiRoller™ plating beads to spread evenly.
- Incubate the plates for overnight at 37 °C.
Electroportation settings
Mode: Exponential protocol
Voltage (V): 1,800 V
Capacitance: 25 uFD
Resistance: 200 Ohms
Cuvette: 1 mm
