ATCC15834 Chemically Competent Agrobacterium cells are made from a specific strain of Rhizobium rhizogenes (formerly Agrobacterium rhizogenes), Agrobacterium rhizogenes pRi15834 (agropine type). Agrobacterium rhizogenes is a soil-borne gram-negative bacterium that can infect most dicotyledons, a few monocotyledons and some gymnosperms. ATCC15834 Chemically Competent Agrobacterium are optimized for the highest transformation efficiencies and are useful for transgenic operations of legumes, tobacco, variety of grasses and other plants. ATCC15834 Agrobacterium rhizogenes strain contains pRi15834 agrobacterium-type Ri plasmid and displays resistance to Rifampicin.
Specifications:
Competent cell type: Chemically Competent
Species: R. rhizogenes
Strain: ATCC15834
Format: Tubes
Transformation efficiency: ≥ 1 x 105 cfu/µg pCAMBIA1391z DNA
Blue/white screening: No
Shipping condition: Dry ice
Reagents Included:
ATCC15834 Chemically Competent Agrobacterium
DNA (pCAMBIA1391z, 500 pg/µl)
Recovery medium
Note: Liquid nitrogen is required.
Storage:
ATCC15834 Chemically Competent Agrobacterium: -80ºC
pCAMBIA1391z control DNA: -20ºC
Recovery medium: 4ºC
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 105 CFU/µg pCAMBIA1391z DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines:
Follow these guidelines when using ATCC15834 Chemically Competent Agrobacterium 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.
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