GoldBio’s AGL-1 Agrobacterium chemically competent cells allow you to obtain high transformation efficiency in applications such as gDNA or cDNA library construction. Our AGL-1 strain harbors the C58 chromosomal backbone with an insertion mutation in its recA recombination gene. This mutation stabilizes recombinant plasmids. AGL-1 also has rifampicin and carbenicillin resistance genes in the genome useful for selection. A functional T-DNA binary system can be built using our AGL-1 strains as the T-DNA region has been deleted from the Ti plasmid pTiBO542 and instead it has a binary vector containing the missing T-region. The binary system makes possible to transfer genetic material into a host plant S genome. Therefore, our system is often used for Agrobacterium-mediated transformation in mono and dicotyledonous species such as Arabidopsis thaliana, maize, and other plants.

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

Product SpecificationsCompetent cell type: Chemical CompetentSpecies: A. tumefaciensStrain: AGL-1Transformation efficiency: ≥4 x 10⁴ cfu/µg pCAMBIA1391z DNABlue/white screening: No

Storage/Handling: This product may be shipped on dry ice. GV3101 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.

• AGL-1Chemically Competent Agrobacterium: 50µl
• DNA (pCAMBIA1391z, 10 ng/µl): 5 µl
• Recovery medium: 1 ml

Quality ControlTransformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥4 x 10⁴ 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 EfficiencyTransformation 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 = 250µg of DNA = 0.00001 Dilution = 10/1000 x 50/1000 = 0.0005 TE = 250/0.00001/0.0005 = 5.0 × 10¹⁰

IPTG, X-Gal and X-Gluc

©蚂蚁淘

没错！最开始，Gold Biotechnology只生产这三个产品，30年后的今天，Goldbio的产品已经超过3000种，涵盖：

琼脂糖树脂

抗生素

生化试剂

缓冲液

克隆与诱导

培养基添加剂

洗涤剂

DNA蛋白质电泳

酶、抑制剂和底物

生长因子

植物研究

蛋白质表达与纯化

还原剂……

目前Goldbio最核心的产品是：荧光素钾盐，X-gluc， 双丙氨磷，草胺磷 和 X-α-gal