AGL1 Electrocompetent Agrobacterium

GoldBio’s AGL-1 Agrobacterium Electrocompetent Cells are optimized to supply the best transformation effectivity, and are perfect for functions requiring excessive transformation efficiencies reminiscent of cDNA or gDNA library building. The AGL-1 pressure has a C58 chromosomal background that carries an insertion mutation in its recA recombination gene which stabilizes recombinant plasmids. It additionally carries rifampicin and carbenicillin resistance in its genome for choice.

AGL-1 accommodates the Ti plasmid pTiBo542 from which the T-DNA area sequences have been deleted. Transformation with a binary vector containing the lacking T-region ends in a practical T-DNA binary system that enables for switch of genetic materials into a bunch plant’s genome. Therefore, this technique is commonly used for Agrobacterium-mediated transformation of Arabidopsis thaliana in addition to maize and different monocots Intact Genomics.

Table 1: Antibiotic disc sensitivity for GoldBio’s Agrobacterium strains (utilizing normal BD antibiotic discs)

	Antibiotic Selection
	Amp	Carb	Chlor		Gent	Kan	Rif	Spect	Strep	Tet
	100 µg/ml	100 µg/ml	30 µg/ml	100 µg/ml	30 µg/ml	50 µg/ml	25 µg/ml	50 µg/ml	50 µg/ml	50 µg/ml
GV3101	I	R	R	PR	R	S	R	S	R	S
EHA 105	R	R/S	R	n/a	R/S	S	R	S	R	S
LBA 4404	S	S	S	n/a	S	S	R	S	R	S
AGL-1	R	R	R	n/a	R	S	R	S	R	S
C58C1	R	R	R	n/a	R	S	R	S	R	S
S = Sensitive R = Resistant R/S = intermediate zones utilizing normal discs. I = development in inhibitory zone with normal disc. “Opaque”, not clear zone of inhibition.

Product Specifications
Competent cell kind: ElectroCompetent
Species: A. tumefaciens
Strain: AGL-1
Format: Tubes
Transformation effectivity: ≥1 x 10⁷ cfu/µg pCAMBIA1391z DNA
Blue/white screening: No

Storage/Handling: This product could also be shipped on dry ice. AGL-1 Agrobacterium Electrocompetent cells needs to be saved at -80°C, pCAMBIA1391z Control DNA needs to be saved at -20°C and restoration medium needs to be saved at 4°C instantly upon arrival. When saved beneath the really useful situations and dealt with appropriately, these merchandise needs to be steady for no less than 1 yr from the date of receipt.

Genomic Features

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

Reagents Needed for One Reaction

• AGL-1 ElectroCompetent Agrobacterium: 25 µl
• DNA (pCAMBIA1391z, 500 pg/µl): 1 µl
• Recovery medium: 1 ml

Quality Control
Transformation effectivity is examined by utilizing the pCAMBIA1391z management DNA provided with the equipment and utilizing the protocol given beneath. Transformation effectivity needs to be ≥6 x 10⁷ CFU/µg pCAMBIA1391z DNA. Untransformed cells are examined for applicable antibiotic sensitivity.

General Guidelines

• Handle competent cells gently as they’re extremely delicate to modifications in temperature or mechanical lysis attributable to pipetting.
• Thaw competent cells on ice, and remodel cells instantly following thawing. After including DNA, combine by tapping the tube gently. Do not combine cells by pipetting or vortexing.

Note: A high-voltage electroporation equipment able to producing discipline strengths of 16 kV/cm is required.

Calculation of Transformation Efficiency

Transformation Efficiency (TE) is outlined because the variety of colony forming items (cfu) produced by remodeling 1 µg of plasmid right into a given quantity of competent cells.

• • TE = Colonies/µg/Dilution
    • Colonies = the variety of colonies counted
    • µg = quantity of DNA reworked in µg
    • Dilution = complete dilution of the DNA earlier than plating

Example: Transform 1 µl of (10 pg/µl) management 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 subsequent day. If you depend 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 × 10^10.