K599 Chemically Competent Agrobacterium rhizogenes (str R) pRi2659 (agropine type) - 18 x 50 µl

This K599 Agrobacterium Chemically competent 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.  K599 Chemically Competent Agrobacterium are optimized for the highest transformation efficiencies and are useful for transgenic operations of corn, soybean (wild 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: Chemically Competent

Species: A. rhizogenes

Strain: K599

Format: Tubes

Transformation efficiency:  ≥ 1 x 104  cfu/µg pCAMBIA1391z DNA

Blue/white screening: No

Shipping condition: Dry ice

K599 Agrobacterium Chemically Competent Cells

DNA (pCAMBIA1391z , 500 pg/µl)

Recovery medium

Note: Liquid nitrogen is required.

K599 Agrobacterium Chemically Competent Cells: -80 ºC

pCAMBIA1391z control DNA: -20 ºC

Recovery medium: 4 ºC

Dry ice

Quality Control

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 104 CFU/µg pCAMBIA1391z Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines

Follow these guidelines when using K599 Agrobacterium Chemically Competent 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.

Example Calculation of Transformation Efficiency

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

TE = Colonies/µg/Plated

Transform 1 µl of (500 pg/µl) pCAMBIA1391z control plasmid into 50 µl of cells, add 950 µl of Recovery Medium. Recover for 3 hours and plate 100 µl. Count the colonies on the plate in two days. If you count 5 colonies, the TE is calculated as follows:

Colonies = 5

µg of DNA = 0.0005

Dilution = 100/1000 = 0.1

TE = 5/.0005/.1 = 1×105