C58C1 (pSuperAgro™ v4) Agrobacterium Electrocompetent Cells

GoldBio’s C58C1 ElectroCompetent 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.

pSuperAgro™ v4 was designed to include AcdS and GabT activity, driven by a single lac
promotor, which both suppresses ethylene evolution and degrades
gamma-aminobutyric acid (GABA) during co-cultivation. The combined
suppression of ethylene and reduction of GABA significantly increases
T-DNA transfer and increases transient and stable transformation
frequencies in both tomato and grass plants.

These
products are sold under license by GoldBio
and require a signed
agreement before fulfilment. The purchase of this product includes a
1-year subscription to use pSuperAgro™ in your research. See license
details below in the technical documentation.

Kit Components

• Competent Cells
• 1 x 12 mL Recovery Media
• 1 x 10 µL Control Plasmid (pCAMBIA1391z Control, 500 pg/µL)

Product Specifications

Competent cell type: ElectroCompetent

Species: A. tumefaciens

Strain: C58C1 (pSuperAgro™ v4)
Transformation efficiency: ≥1 x 10⁷ cfu/µg pCAMBIA1391z DNA

Blue/white screening: No

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

• ≥1 x 10⁷ cfu/µg efficiency with electroporation.

Reagents Needed for One Reaction

• C58C1 (pSuperAgro™ v4) ElectroCompetent Agrobacterium: 25 µL
• DNA (pCAMBIA1391z Control, 500 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 ≥1 x 10
⁷ 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

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