GV2260 Agrobacterium Electrocompetent Cells

GV2260 Agrobacterium Electrocompetent cells are a derivative of C58 Agrobacterium. GV2260 has the atu3300 gene in its linear chromosome and is a non-oncogenic Ti plasmid from which the T-region, including the entire TL- and TR-region, has been deleted and substituted by sequences derived from pBR322. pGV2260 is used as acceptor Ti plasmid and includes both carbenicillin and rifampicin resistance genes.

Kit Components

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

Product Specifications
Competent cell type: Electrocompetent
Species: A. tumefaciens
Strain: C58
Format: Tubes
Transformation efficiency: ≥1 x 10⁶ cfu/µg pCAMBIA1391z DNA
Blue/white screening: No

Storage/Handling
This product may be shipped on dry ice. GV2260 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.

Reagents Needed for One Reaction

• GV2260 Chemically Competent Agrobacterium: 50 µL
• DNA (pCAMBIA1391z, 10 ng/µL): 5 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 ≥1 x 10⁶ 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