GoldBio’s AGL-1 Agrobacterium Electrocompetent Cells are optimized to produce the highest transformation efficiency, and are ideal for applications requiring high transformation efficiencies such as cDNA or gDNA library construction. The AGL-1 strain has a C58 chromosomal background that carries an insertion mutation in its recA recombination gene which stabilizes recombinant plasmids. It also carries rifampicin and carbenicillin resistance in its genome for selection. AGL-1 contains the Ti plasmid pTiBo542 from which the T-DNA region sequences have been deleted. Transformation with a binary vector containing the missing T-region results in a functional T-DNA binary system that allows for transfer of genetic material into a host plant’s genome. Therefore, this system is often used for Agrobacterium-mediated transformation of Arabidopsis thaliana as well as maize and other monocots.
Competent cell type: ElectroCompetent
Species: A. tumefaciens
Transformation efficiency: ≥1 x 107 cfu/µg pCAMBIA1391z DNA
Blue/white screening: No
Storage/Handling: This product may be shipped on dry ice. AGL-1 Agrobacterium Electrocompetent cells should be stored at -80°C, pCAMBIA1391z 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.
Reagents Needed for One Reaction
Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥6 x 107 CFU/µg pCAMBIA1391z DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
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.
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