K599 ElectroCompetent Agrobacterium rhizogenes (str R) pRi2659 (agropine type) - 6 x 50 µl

This K599 Agrobacterium ElectroCompetent Cells are made from a specific strain of Agrobacterium rhizogenes, Agrobacterium rhizogenes (str R) pRi2659 (agropine type). K599 was originally isolated from cucumber exhibiting hairy root disease symptoms and has been widely used for hairy root transformation.

Agrobacterium rhizogenes is a soil-borne gram-negative bacterium that can infect most dicotyledons, a few monocotyledons, and some gymnosperms.  IG’s K599 Agrobacterium ElectroCompetent Cells is optimized for the highest transformation efficiencies and is useful for transgenic operations of corn, soybean, cotton, peanut, dandelion, cowpea, and other plants.  K599 Agrobacterium rhizogenes strain contains pRi2659 agrobacterium-type Ri plasmid and displays Spectinomycin resistance.

Competent cell type: Electrocompetent

Species: A. rhizogenes

Strain: K599

Format: Tubes

Transformation efficiency: ≥ 1 x 105  cfu/µg pCAMBIA1391z DNA

Blue/white screening: No

Shipping condition: Dry ice

K599 Agrobacterium ElectroCompetent Cells: 25 µl

DNA (pCAMBIA1391z , 500 pg/µl): 1 µl

Recovery medium: 1 ml

K599 Agrobacterium ElectroCompetent Cells: -80 ºC

pCAMBIA1391z control DNA: -20 ºC

Recovery medium: 4 ºC

Dry ice

Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol in this manual. Transformation efficiency should be ≥1 x 105 CFU/µg pCAMBIA1391z DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

Follow these guidelines when using K599 Agrobacterium ElectroCompetent Cells:

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 such as Bio-Rad Gene Pulser II #165-2105, capable of generating field strengths of 16 kV/cm is required.

Transformation Efficiency (TE) is defined as the number of colony-forming units (cfu) produced by transforming 1µg of the plasmid into a given volume of competent cells.

TE = Colonies/µg/Plated

Transform 1 µl of (500 pg/µl) pCAMBIA1391z control plasmid into 25 µl of cells and add 974 µl of Recovery Medium. Recover for 3 hours and plate 100 µl. Count the colonies on the plate in two days. If you count 500 colonies, the TE is calculated as follows:

Colonies = 500

µg of DNA = 0.0005

Dilution = 100/1000 = 0.1

TE = 500/.0005/.1 = 1×107