Product overview Technical/Spec Support/Documents FAQs Tips for usage Related products General informationChampion™ Competent Cells are chemically competent cells, which were prepared by SMOBIO to make E. coli perform excellent transformation efficiency. Standard transformation protocol is recommended for large plasmids or non-ampicillin selection. Time-saving transformation protocol is recommended for simple and rapid transformation. Champion™ Competent Cells are one of the fastest and simplest ready-to-use competent cell products in the world.Kit contentsChampion™ Competent CellspUC19 Control Plasmid (5 μl, 10-4 μg/μl)Champion™ Transformation Protocol CardShipping conditionThroughout the shipping process, the temperature is maintained under -70°C. Storage and expiration Champion™ Competent Cells must be stored between -70°C to -80°C. Subsequent freeze-thaw cycles will reduce transformation efficiency. If high efficiency is required for the experiment, do not use aliquots that have gone through several freeze-thaw cycles. The efficiency of Champion™ Competent Cells lasts for 1 year with proper storage. Genotypes and applicationsProduct NameGenotypeApplicationChampion™ 109 Highe14(McrA-)recA1 endA1 gyrA96 hsdR17(rk-, mK+) phoA supE44 thi-1 relA1 ∆(lac-proAB) (F’traD36 proAB lacIqZ∆M15Appropriate for blue-white color and robotic screening. It is a fast growing strain forming visible colonies within 8~10 hours.Champion™ 21F’ ompT hsdSβ(rβ-mβ-) dcm gal λ (DE3)Appropriate host for recombinant protein expression using T7-based expression vectors.Champion™ DH5α HighrecA1 endA1 gyrA96 hsdR17(rk-, mK+) phoA supE44 relA1 thi-1 ∆(lacZYA-argF)U 169 φ80 ∆(lacZ)M15 F’Suitable for cloning with large plasmid and cDNA library construction, and also for blue-white colony selection.Items and ordering informationProduct NameCompatible to E. coli strainEfficiency (cfu/μg)QuantityCat. No.Champion™ 109 HighE. coli JM109 >1 x 108100 μl x 80 vialsCC0202100 μl x 24 vialsCC0204Champion™ 21E. coli BL21 (DE3) >1 x 107100 μl x 80 vialsCC2102100 μl x 24 vialsCC2104Champion™ DH5α HighE. coli DH5α >3 x 108100 μl x 80 vialsCC5202100 μl x 24 vialsCC5204  ManualManual_Champion™ Competent Cell SDSSDS_Champion™ Competent CellFlyerChampion™ Competent CellProtocol card Will transformation efficiency be reduced when Champion™ competent cells are thawed, dispensed and refrozen repeatedly?When Champion™ competent cells are thawed dispensed in aliquots and refrozen within 1 min, the transformation efficiency will be 10~20% reduced compared to the first time use.What is the advantage of thawing competent cells with circulating water instead of still water?Using circulating water can reduce the thawing time. Less thawing time shows higher efficiency than long thawing time. Is there any difference of transformation efficiency between using plating beads, bending glass rod and plating loop? The bending glass rod shows best efficiency and the plating loop show less efficiency than other method. For handle a lot samples at the same time the plating beads are recommended. Is 1 second vortex critical for mixture the DNA?One second vortex provides more reliable transformation efficiency (1.1 times compare with mixed by finger tapping)Will heat shock affect the transformation efficiency?  Heat shock treatment will enhance the efficiency about 1~2X versus non-heat shock method. Factors Affecting Transformation Efficiency Thawing methods Shorter thawing time is more efficient than a longer thawing time. Slow thawing caused by power shortages and unstable freezers will result in decreased efficiency. Size of plasmid Plasmid size affect the efficiency greatly. The efficiency of transforming a supercoiled 2.7 kb is approximately 100 times higher than that of a 10 Kb plasmid (using time-saving transformation protocol). For large plasmids (> 6 kb), standard transformation protocol is recommended.Heat shock treatmentHeat shock treatment will enhance the efficiency about 1~2 folds versus non-heat shock method. Plating methods Bent glass rods show the greatest efficiency, while plating loop shows less efficiency than plating beads. When handling a large quantity of samples at the same time, plating beads are recommended. Concentration of antibiotic Antibiotic concentration is critical to use of the time-saving transformation protocol. AntibioticConcentrationAmpicillin (Ap)20 μg/mlKanamycin (Km)25 μg/mlTetracycline (Tc)7.5 μg/mlChloramphenicol (Cm)20 μg/mlFor plasmid size <6 Kb, the efficiency of kanamycin selection is usually 3~10 times less than the ampicillin selection. For plasmid size > 6 Kb, the efficiency of kanamycin selection is much lower than ampicillin. We suggest using the standard transformation protocol (with heat shock and recovery steps) to enhance the efficiency.Product NameCompatible to E. coli strainEfficiency (cfu/μg)QuantityCat. No.Champion™ 109 HighE. coli JM109 >1 x 108100 μl x 80 vialsCC0202100 μl x 24 vialsCC0204Champion™ 21E. coli BL21 (DE3) >1 x 107100 μl x 80 vialsCC2102100 μl x 24 vialsCC2104Champion™ DH5α HighE. coli DH5α >3 x 108100 μl x 80 vialsCC5202100 μl x 24 vialsCC5204 High Fidelity PCR amplificationAmplification of target gene with HiFi™ DNA polymerase to minimize error rate.[TF1000] SMO-HiFi™ DNA Polymerase, (1 U/μl, 100 U)[TF3000] G-HiFi™ DNA Polymerase, (1 U/μl, 100 U)Gel electrophoresisStaining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency.   Safe fluorescent dyes[NS1000] FluoroVue™ Nucleic Acid Gel Stain (10,000X), 500 μl[DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl[DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 mlBlue-light illuminator[VE0100] B-BOX™ Blue Light LED Epi-illuminatorLigationBlund-end PCR amplicons can directly ligate with PCR cloning vector.  [CV1000] GetClone™ PCR Cloning Vector, 20 Rxn[CV1100] GetClone™ PCR Cloning Vector II, 20 RxnColony PCRAnalyze colonies with PCR master mix to save preparation time.  [TP1100] ExcelTaq™ 5X PCR Master Mix, 200 Rxn[TP1200] ExcelTaq™ 5X PCR Master Dye Mix, 200 Rxn