AGL-1 Agrobacterium Electrocompetent Cells

Product Description

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.

Product Specifications
Competent cell type: ElectroCompetent
Species: A. tumefaciens
Strain: AGL-1
Format: Tubes
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.

Genomic Features

  • ≥5 x 107 cfu/µg efficiency with electroporation.

Reagents Needed for One Reaction

  • GV3101 ElectroCompetent Agrobacterium: 25 µl
  • DNA (pCAMBIA1391z, 100 pg/µl): 1 µl
  • Recovery medium: 1 ml

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 ≥6 x 107 CFU/µg pCAMBIA1391z 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

Transform 1 µl of (100 pg/µl) pCAMBIA1391z control plasmid into 25 µl of cells, add 975 µl of Recovery Medium. Recover for 3 hours and plate 100 µl. Count the colonies on the plate in two days. If you count 500 colonies, the TE is calculated as follows:

Colonies = 500
µg of DNA = 0.0005
Dilution = 100/1000 = 0.1
TE = 500/0.0005/0.1 = 1.0 × 107

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