Introduction to Competent Cells
What are competent cells?
Cell competence refers to a cell’s ability to take up foreign (extracell...
Hub page housing all our competent cells, grouped.
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GoldBio offers an extensive collection of efficient chemically competent and electrocompetent cells with high transformation efficiencies to match your specific downstream applications: general cloning, protein expression, library construction, and many more.
- GoldBio's wide range of chemically competent and electrocompetent cells are optimized to provide lot-to-lot consistency and up to 10X higher transformation efficiency.
- Our enhanced recovery medium (SOC) formulation accompanies all of GoldBio's competent cells.
- We carry multiple Endonuclease I-deficient (endA1) competent bacterial cell strains to suit your cloning needs and ensure your transformations result in high quality and yield of plasmid DNA.
- Choose from E. coli strains with different selectable markers and screening options including the classic blue/white selection.
- GoldBio cells are ideal for distinct downstream applications from routine cloning/subcloning and protein expression to the more complex library construction, and cloning of difficult targets.
- Our electrocompetent Agrobacterium cells contain an efficient T-DNA binary system for genetic material transfer into the host plant cells.
- GoldBio's amber suppressor (SupE) and non-amber suppressor (MC1061F\’) strains are ideal for phage display library screening.
CHOOSE THE BEST COMPETENT CELLS FOR YOUR APPLICATION!
Our DH5-alpha, BL21, BL21(DE3), DH10B, DH10B-Pro, DL39(DE3), Agrobacterium, L. lactis, SS320 and TG1 Phage Display cells are available in multiple formats to provide you with flexibility when planning your experiments. GoldBio also provides easy-to-follow protocols, FAQs, protocol videos and selection guides to ensure success of your transformations!
GoldBio Competent Cells Consistently Exhibit Higher Transformation Efficiencies
DH10B chemically competent E. coli cells and DH10B Electrocompetent E. coli cells exhibit higher transformation efficiency than competitors. Each transformation with DH10B chemically competent E. coli cells was performed using 1 ml of final volume including recovery media, plating 50 μl after a 10 fold-dilution (left). For electrocompetent cell comparison, DH10B Electrocompetent E. coli cells were electroporated using a 1 ml of final volume including recovery media, plating 50 μl after 100-fold dilution (right).
BROWSE OUR TRANSFORMATION AND COMPETENT CELL RESOURCES
Your time in the lab is valuable! In this section you will find GoldBio's transformation tools designed to help support and simplify your competent cell transformations. You can access specific chemical transformation or electroporation protocols for each of our competent cell lines, antibiotic information, colony screening protocols, instructional blog articles, detailed guides containing competent cells characteristics and downstream applications, FAQs, electroporation troubleshooting tips and video protocols.
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Electroporation is a technique commonly used in transformation assays to introduce DNA into an electrocompetent cell. During electroporation a transient current or electrical pulse is applied to cells that have been suspended in a solution. The current momentarily increases the permeability of the cell's membrane, creating holes (a pathway) that allow DNA (plasmids) to pass through and enter the cell. Generally, electroporation can be more effective than chemical transformation.
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Product Description
GoldBio’s BL21 (DE3) Electrocompetent E. coli cells are suitable for high efficiency transformation and routine protein expression. Increased cloning efficiencies versus typical BL21 cells makes the BL21 (DE3) Electrocompetent E. coli cells ideal for construction of complex expression libraries and feature ≥1 x 107 cfu/µg efficiency with electroporation. BL21 (DE3) Electrocompetent E. coli cells have a widely used background, feature a T7 expression strain, are deficient in both lon (1) and ompT proteases. These cells are resistant to phage T1(fhuA2) and are B strain.
Product Specifications
Competent cell type: ElectroCompetent
Derivative of: BL21 (DE3)
Species: E. coli
Format: Tubes
Transformation efficiency: ≥1 x 107 cfu/µg pUC19 DNA
Blue/white screening: Yes
Storage/Handling: This product may be shipped on dry ice. BL21 (DE3) Electrocompetent E. coli cells should be stored at -80°C, pUC19 Control DNA should be stored at -20°C and recovery medium should be stored at 4°C immediately upon arrival. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.
Genomic Features
- ≥1 x 107 cfu/µg efficiency with electroporation
- Widely used host background
- T7 Expression Strain
- Deficient in both lon (1) and ompT proteases
- Resistant to phage T1 (fhuA2)
- B Strain
Genotype
F–ompT hsdS(rB– mB–) gal dcm λ(DE3)
Reagents Needed for One Reaction
- BL21 (DE3) electrocompetent cells: 25 µl
- DNA (or pUC19 Control, 10 pg/µl): 1 µl
- Recovery medium: 1 ml
Quality Control
Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 107 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
- 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.
Note: A high-voltage electroporation apparatus capable of generating field strengths of 16 kV/cm is required.
Calculation of Transformation Efficiency
Transformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1 µg of plasmid into a given volume of competent cells.
- TE = Colonies/µg/Dilution
- Colonies = the number of colonies counted
- µg = amount of DNA transformed in µg
- Dilution = total dilution of the DNA before plating
Example: Transform 1 µl of (10 pg/µl) control plasmid into 25 µl of cells, add 975 µl of Recovery Medium. Dilute 10 µl of this in 990 µl of Recovery Medium and plate 50 µl. Count the colonies on the plate the next day. If you count 250 colonies, the TE is calculated as follows:
Colonies = 250
µg of DNA = 0.00001
Dilution = 10/1000 x 50/1000 = 0.0005
TE = 250/0.00001/0.0005 = 5.0 × 1010
Technical Documentation
P
Protocol
O
Other
C
Certificate of Analysis
S
Safety Data Sheet
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