C58C1 Agrobacterium Chemically Competent Cells

GoldBio’s C58C1 Chemically Competent Agrobacterium cells allow you to obtain high transformation efficiency in applications such as gDNA or cDNA library construction. C58C1 has the chromosomal backbone of C58 but it has been cured of the Ti plasmid pTiC58. Our C58C1 strain harbors streptomycin and rifampicin resistance genes. C58C1 Competent cells are optimized for genetic transformation of plants such as Arabidopsis but can be used in many other plants.

Kit Components

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

Competent cell type: Chemical Competent
Species: A. tumefaciens
Strain: C58C1
Transformation efficiency: ≥1 x 10⁵ cfu/µg pCAMBIA1391z DNA
Blue/white screening: No

Storage/Handling: This product may be shipped on dry ice. C58C1 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

• C58C1 Chemical Competent Agrobacterium: 50 µl
• DNA (pCAMBIA1391z, 10 ng/µl): 5 µl
• Recovery medium: 1 ml

Table 1: Antibiotic disc sensitivity for GoldBio’s 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 x 10⁵ 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