EHA105 (pSoup) Agrobacterium Chemically Competent Cells

GoldBio’s EHA105 (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, chloramphenicol, and streptomycin, providing versatility in selection and antibiotic resistance EHA105 strain contains a rifampicin resistance gene (rif) and an amber basic Ti plasmid pEHA105 (pTiBo542DT-DNA) without self-transport function, containing the vir gene.

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 EHA105 (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.

GoldBio’s EHA105 Agrobacteriumstrain was generated, and primary clone supplied by Dr. Elizabeth Hood.

Table 1: Antibiotic disc sensitivity for GoldBio’s EHA105 Agrobacterium strains (using standard BD antibiotic discs)

Product Specifications

Competent cell type: Chemical Competent

Species: A. tumefaciens

Strain: EHA105 (pSoup)
Transformation efficiency: ≥5 x 10³ cfu/µg pCAMBIA1391z DNA

Blue/white screening: No

Storage/Handling: This product may be shipped on dry ice. EHA105 (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

• EHA105 (pSoup) Chemical 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 ≥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