DH10B Competent Cells - FAQs

View additional product information for DH10B Competent Cells - FAQs (EC0113)

4 product FAQs found

What is the storage media for the competent cells?

Preparation procedures and formulations for all of our competent cells are proprietary. All chemically competent cells are delivered in an aqueous solution that contains a mixture of salts, along with a freezing stabilizer such as glycerol or DMSO.

I need a competent cell line that will generate as much DNA as possible from my vector of interest. What do you recommend?

The three most important things in this case are: endA- genotype, a high copy number origin of replication, and the culture scale. In addition, make sure (a) the culture is grown at 37 degrees C (if lower, the copy number can be lower), (b) try a super-rich media, like Terrific broth, (c) aerate the culture well (the volume of media should be no more than 10% of the volume of the flask), and (d) shake at 200-250 rpm.

What is the formulation of the SOC medium that is provided with competent cells?

SOC (Super Optimal Catabolite) Medium Preparation (for 1 Liter):

1) To a 2 Liter flask with stir bar add the following:
- Bacto Tryptone 20 g
- Yeast Extract 5 g
- Sodium Chloride (NaCl) 0.58 g
- Potassium Chloride (KCl) 0.186 g
2) Add sterile water to a final volume of 1 Liter.
3) Mix well on magnetic stir plate for 5-10 minutes or until all of the ingredients are well mixed and completely dissolved.
4) Autoclave 30 minutes.
5) Allow to cool to room temperature.
6) Add 10 ml of sterile 2M Magnesium Solution (1M Magnesium sulfate, 1M Magnesium chloride)and mix well.
7) Add 10 ml of sterile 2M Glucose and mix well. (Final Glucose concentration is 20 mM).

How can I obtain the highest possible transformation efficiency with Thermo Scientific DH5α and DH10B competent cells?

To achieve the highest possible transformation efficiency with Thermo Scientific DH5α (Cat. No. EC0112) and DH10B (Cat. No. EC0113) competent cells, please follow the transformation protocol provided with the product and pay attention to the following details:

- Make sure to use wet well-pressed ice to maximize cold surface volume in all the transformation steps.
- Perform the steps quickly and accurately.
- Use chilled microcentrifuge tubes and bury the whole tube up to the cap in the wet ice.
- Do not exceed the recommended heat shock time and transfer the tube with cells back to the wet ice immediately.