GoldBio’s C58C1 Chemically Competent Agrobacteriumcells allow you to obtain high transformation efficiency in applications such as gDNA or cDNA library construction. C58C1 has the chromosomal backbone of C58 but it has been cured of the Ti plasmid pTiC58. Our C58C1 strain harbors streptomycin and rifampicin resistance genes. C58C1 Competent cells are optimized for genetic transformation of plants such as Arabidopsis but can be used in many other plants.
Table 1: Antibiotic disc sensitivity for GoldBio’s Agrobacterium strains (using standard 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 | S | R | S | R | S |
C58C1 | R | R | R | n/a | R/S | S | R | S | R | S |
S = SensitiveR = ResistantR/S =intermediate zones using standard discs.I =growth in inhibitory zone with standard disc. “Opaque”, not clear zone of inhibition. |
Product SpecificationsCompetent cell type: Chemical CompetentSpecies: A. tumefaciensStrain: C58C1Transformation efficiency: ≥1 x 105 cfu/µg pCAMBIA1391z DNABlue/white screening: No
Storage/Handling: This product may be shipped on dry ice. C58C1 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.
Reagents Needed for One Reaction
Quality Control
Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 105 CFU/µg pCAMBIA1391z DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
Calculation of Transformation Efficiency
Transformation 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.
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 = 0.00001Dilution = 10/1000 x 50/1000 = 0.0005TE = 250/0.00001/0.0005 = 5.0 × 1010
IPTG, X-Gal and X-Gluc
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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