Introduction to Competent Cells
What are competent cells? Cell competence refers to a cell’s ability to take up foreign (extracell...
GoldBio’s GB10B™ Chemically Competent E. coli cells are high efficiency cells, suitable for a wide variety of applications such as cloning and sub-cloning. GB10B™ Chemically Competent E. coli cells have multiple features including the ɸ80lacZ∆M15 marker, which provides α-complementation of the β-galactosidase gene with blue/white screening protocol. These cells also have the mcrA genotypic marker and the mcrBC, mrr deletion, which allows for cloning of DNA that contains methylcytosine and methyladenine.
Kit Components
Product Specifications
Competent cell type: Chemically competent
Equivalent to: DH10B
Species: E. coli
Transformation efficiency: ≥8.2 x 106 cfu/µg pUC19 DNA
Blue/white screening: Yes
Storage/Handling: This product may be shipped on dry ice. GB10B™ Chemically Competent 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
Genotype
F – mcrA ∆(mrr-hsdRMS-mcrBC) endA1 recA1 φ80dlacZ∆M15 ∆lacX74 araD139 ∆(ara, leu)7697 galU galK rpsL (StrR) nupG λ-
Reagents Needed for One Reaction
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 ≥8.2 x 106 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
Calculation of Transformation Efficiency
Transformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1µg of plasmid into a given volume of competent cells.
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
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CC-100-5x100 1 week | 5 x 100 μL | $ 84.00 | |
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CC-103-10x50 In stock | 10 x 50 μL | $ 84.00 | |
CC-103-15x50 In stock | 15 x 50 μL |
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CC-103-20x50 In stock | 20 x 50 μL |
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