Intact Genomics (ig®) Methionine-Auxotrophic LBA4404Met Chemically Competent Agrobacterium cells include modifications so that they will not grow unless free methionine is added to Minimal medium. This prevents the bacteria from overgrowing plant tissues when used for plant transformation. Methionine- Auxotrophic LBA4404Met Chemically Competent Agrobacterium cells are optimized for the highest transformation efficiencies. The LBA4404Met strain is useful for transgenic operations of tomatoes, tobacco and other plants. LBA4404Met contains a rifampicin resistance gene (rif). LBA4404Met strain also contains a octoprine-type Ti plasmid pAL4404Met without self transport function, which contains the vir gene.
Custom Aliquots Available
Benefits
- Methionine Auxotrophic
- Enables development of more efficient transformation systems
- Reduced bacterial overgrowth during co-cultivation
- Decreased need for antibiotics
Specifications
Competent cell type: Chemically CompetentSpecies: A. tumefaciensFormat: TubesTransformation efficiency: ≥ 1 x 105 cfu/µg pUC19 DNAShipping condition: Dry ice
Reagents Included
- ig® Methionine-Auxotrophic LBA4404Met Chemically Competent Agrobacterium Cells
- DNA (pCAMBIA1391z, 500 pg/µl)
- Recovery medium
Storage
- ig® Methionine Auxotrophic LBA4404Met Chemically Competent Agrobacterium Cells: -80 ºC
- pCAMBIA1391z control DNA: -20 ºC
- Recovery medium: 4 ºC
Quality Control
Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. 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 ig® Methionine-Auxotrophic LBA4404Met Chemically Competent Agrobacterium :
- 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.
Example Calculation of Transformation EfficiencyTransformation 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.
TE = Colonies/µg/Dilution
Transform 1 µl of (10 pg/µl) pUC19 control plasmid into 50 µl of cells, add 950 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 100 colonies, the TE is calculated as follows:
Colonies = 5µg of DNA = 0.0005Dilution = 100/1000=0.1TE = 5/.0005/.1 = 1.0×1051076-05 1076-15
