Intact Genomics Agrobacterium tumefaciens AGL1 (AGL-1) cells are optimized for the highest transformation efficiencies which are ideal for applications requiring high transformation efficiencies, such as with cDNA or gDNA library construction.
The AGL1 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. AGL1 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 the transfer of genetic material into a host plant’s genome. Therefore, this system is often used for the Agrobacterium-mediated transformation of Arabidopsis thaliana as well as maize and other monocots.
SpecificationsCompetent cell type: Chemically CompetentSpecies: A. tumefaciensStrain: AGL1Format: TubesTransformation efficiency: ≥ 1 x 105 cfu/µg pCAMBIA1391z DNABlue/white screening: NoShipping condition: Dry ice
Reagents Included
- AGL1 Chemically Competent Agrobacterium
- DNA (pCAMBIA1391z, 500 pg/µl)
- Recovery medium
Note: Liquid nitrogen is required.
StorageAGL1 Chemically Competent Agrobacterium: -80 ºCpCAMBIA1391z control DNA: -20 ºCRecovery medium: 4 ºC
Quality Control
Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol in this manual. Transformation efficiency should be ≥1 x 105 CFU/µg pCAMBIA1391z DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
Follow these guidelines when using AGL1 Chemically Competent Agrobacterium cells:
- 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.
Calculation of Transformation EfficiencyTransformation Efficiency (TE) is defined as the number of colony-forming units (cfu) produced by transforming 1µg of the plasmid into a given volume of competent cells.
TE = Colonies/µg/Plated
Transform 1 µl of (500 pg/µl) pCAMBIA1391z control plasmid into 50 µl of cells, and add 950 µl of Recovery Medium. Recover for 3 hours and plate 100 µl. Count the colonies on the plate in two days. If you count 5 colonies, the TE is calculated as follows:
Colonies = 5µg of DNA = 0.0005Dilution = 100/1000 = 0.1TE = 5/.0005/.1 = 1×105
Please note, all agrobacterial strains are not well-studied for antibiotic resistance and there are many agrobacterial strains. Therefore, it is the customer’s responsibility to make sure his/her vectors are compatible with the Agrobacterial strains if he/she uses an alternate antibiotic selection than kanamycin-selection.1082-06 1082-18 1083-10
