GV3101 (pSoup) Agrobacterium Chemically Competent Cells

GoldBio’s GV3101 Agrobacterium chemically competent cells are a powerful tool for high efficiency transformation of plants. The GV3101 strain is optimized for cDNA and gDNA library construction, and possesses resistance to rifampicin, gentamicin, and tetracycline. The T-DNA region of the GV3101 Ti plasmid has been deleted, but restoration of the binary vector T-region can fully enable T-DNA binary system function.

The presence of the pSoup helper plasmid is required for the replication of pGreen, 62SK, and pGs2 series plasmids, and also helps with Agrobacterium-mediated transformation of dicotyledonous plants like Arabidopsis thaliana, tobacco, and potato, as well as monocotyledonous plants such as corn.

Kit Components

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

Competent cell type: Chemically Competent
Species: A. tumefaciens
Strain: GV3101 (pSoup)
Transformation efficiency: ≥1.5 x 10³ cfu/µg pCAMBIA1391z DNA
Blue/white screening: No

Storage/Handling: This product may be shipped on dry ice. GV3101 (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.

Reagents Needed for One Reaction

• GV3101 (pSoup) chemically competent Agrobacterium: 50 µL
• DNA (pCAMBIA1391z Control, 10 ng/µL): 5 µL
• Recovery medium: 1 mL

Table 1: Antibiotic disc sensitivity for GoldBio’s GV3101 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 ≥1.5 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