Thermo Scientific™

DH5α Competent Cells

Thermo Scientific DH5α Competent Cells are high efficiency, chemically competent E. coli cells ideal for construction of gene banks orRead more

Catalog Number	Quantity
EC0112	10 x 100 μL

Catalog number EC0112

Price (MXN)

Quantity:

10 x 100 μL

Request bulk or custom format

Thermo Scientific DH5α Competent Cells are high efficiency, chemically competent E. coli cells ideal for construction of gene banks or generation of cDNA libraries using plasmid-derived vectors. The φ80dlacZΔM15 marker provides α-complementation of the β-galactosidase gene from pUC or similar vectors to allow blue/white colony screening on bacterial agar plates containing Bluo-Gal or X-Gal. RecA1 and endA1 mutations in DH5α cells improve insert stability and quality of the extracted plasmid DNA.

• High transformation efficiency: >1x10⁹ cfu/μg pUC19 DNA
• Suitable for routine and high-throughput cloning applications
• Genetic markers that allow for blue/white colony screening
• Convenient two reactions per vial packaging
• S.O.C. Outgrowth medium included

Applications
DH5α Competent Cells are suitable for variety of applications requiring high transformation efficiency:
• Efficient DNA cloning derived from PCR, cDNA-generation reactions
• recA1 marker facilitates working with difficult to transform DNA
• Ideal for generation of cDNA libraries using plasmid-derived vectors
• Site-directed mutagenesis

Genotype
F– φ80lacZΔ M15 Δ (lacZYA-argF) U169 recA1 endA1 hsdR17 (rK– mK+) phoA supE44 λ- thi–1 gyrA96 relA1

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Bacterial or Yeast StrainDH5α

Blue/White ScreeningYes

Cell TypeChemically Competent

Cloning Methylated DNANo

Contains F' EpisomeLacks F' Episome

EfficiencyHigh Efficiency (>1x10^9 cfu/μg pUC19 DNA)

For Use With (Application)Cloning

High-throughput CompatibilityNot High-throughput Compatible

LibrariescDNA

Product LineThermo Scientific

Product TypeCompetent Cell

Propagating ccdB VectorsNot for ccdB vector propagation

Quantity10 x 100 μL

Shipping ConditionDry Ice

FormatTube

SpeciesE. coli

Unit SizeEach

Contents & Storage

• 10 x DH5α Competent Cells (100 μL)
• pUC19 DNA (50 μL) (10 pg/μL)
• S.O.C. Medium (5 mL)

Store at -80°C.

Frequently asked questions (FAQs)

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.