AGL-1 (pSoup) Agrobacterium Chemically Competent Cells

GoldBio’s AGL-1 (pSoup) chemically competent Agrobacterium cells are optimized for high transformation efficiencies, making them ideal for applications such as cDNA or gDNA library construction. These cells are resistant to rifampicin, carbenicillin, and streptomycin, providing versatility in selection and antibiotic resistance. The Ti plasmid pTiBo542DT-DNA has been engineered to have the T-DNA region deleted, which can be complemented with a binary vector containing the missing T-region to restore the T-DNA binary system function. This feature enables efficient and precise transformation of plant genomes.

The pSoup plasmid present in these cells is required for the replication of pGreen, 62SK, and pGs2 series plasmids, increasing the range of applications for the AGL-1 (pSoup) system. These competent cells are suitable for Agrobacterium-mediated transformation of dicots such as Arabidopsis thaliana, tobacco, potato, as well as monocots like corn, making them an ideal choice for plant molecular biology research.

Kit Components

• Competent Cells
• 1 x 12 mL Recovery Media
• 1 x 25 µl Control Plasmid (pCAMBIA1391z, 10 ng/µl)

Competent cell type: Chemically Competent
Species: A. tumefaciens
Strain: AGL-1 (pSoup)
Transformation efficiency: ≥3 x 10³ cfu/µg pCAMBIA1391z DNA
Blue/white screening: No

Storage/Handling: This product may be shipped on dry ice. AGL-1 (pSoup) Agrobacterium chemically competent 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.

• AGL-1 (pSoup) chemically competent Agrobacterium: 50 µl
• DNA (pCAMBIA1391z, 10 ng/µl): 5 µl
• Recovery medium: 1 ml

Table 1: Antibiotic disc sensitivity for GoldBio’s AGL-1 Agrobacterium strains (using standard BD antibiotic discs)

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 ≥3 x 10³ 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.

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

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