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General information
Champion™ 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 contents
Champion™ Competent Cells
pUC19 Control Plasmid (5 μl, 10-4 μg/μl)
Champion™ Transformation Protocol Card
Shipping condition
Throughout 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 applications
Product Name | Genotype | Application |
Champion™ 109 High | e14(McrA-)recA1 endA1 gyrA96 hsdR17(rk-, mK+) phoA supE44 thi-1 relA1 ∆(lac-proAB) (F’traD36 proAB lacIqZ∆M15 | Appropriate for blue-white color and robotic screening. It is a fast growing strain forming visible colonies within 8~10 hours. |
Champion™ 21 | F’ ompT hsdSβ(rβ-mβ-) dcm gal λ (DE3) | Appropriate host for recombinant protein expression using T7-based expression vectors. |
Champion™ DH5α High | recA1 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 information
Product Name | Compatible to E. coli strain | Efficiency (cfu/μg) | Quantity | Cat. No. |
Champion™ 109 High | E. coli JM109 | >1 x 108 | 100 μl x 80 vials | CC0202 |
100 μl x 24 vials | CC0204 | |||
Champion™ 21 | E. coli BL21 (DE3) | >1 x 107 | 100 μl x 80 vials | CC2102 |
100 μl x 24 vials | CC2104 | |||
Champion™ DH5α High | E. coli DH5α | >3 x 108 | 100 μl x 80 vials | CC5202 |
100 μl x 24 vials | CC5204 |
Manual
Manual_Champion™ Competent Cell
SDS
SDS_Champion™ Competent Cell
Flyer
Champion™ Competent Cell
Protocol 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 treatment
Heat 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.
Antibiotic | Concentration |
Ampicillin (Ap) | 20 μg/ml |
Kanamycin (Km) | 25 μg/ml |
Tetracycline (Tc) | 7.5 μg/ml |
Chloramphenicol (Cm) | 20 μg/ml |
For 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 Name | Compatible to E. coli strain | Efficiency (cfu/μg) | Quantity | Cat. No. |
Champion™ 109 High | E. coli JM109 | >1 x 108 | 100 μl x 80 vials | CC0202 |
100 μl x 24 vials | CC0204 | |||
Champion™ 21 | E. coli BL21 (DE3) | >1 x 107 | 100 μl x 80 vials | CC2102 |
100 μl x 24 vials | CC2104 | |||
Champion™ DH5α High | E. coli DH5α | >3 x 108 | 100 μl x 80 vials | CC5202 |
100 μl x 24 vials | CC5204 |
High Fidelity PCR amplification
Amplification 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 electrophoresis
Staining 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 ml
Blue-light illuminator
[VE0100] B-BOX™ Blue Light LED Epi-illuminator
Ligation
Blund-end PCR amplicons can directly ligate with PCR cloning vector.
[CV1000] GetClone™ PCR Cloning Vector, 20 Rxn
[CV1100] GetClone™ PCR Cloning Vector II, 20 Rxn
Colony PCR
Analyze 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
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实验室很早就买的这个PCR仪，一直放着没有用，现在想用了，找不到说明啦，求帮助
2、仪器功能：进口和国产仪器主要功能：如保存，过温保护，断电保护，热盖功能等基本上是一样的，只是附带功能上有所差别，比如：进口的显示屏是大型的触摸式荧屏操作，国产的大多是触摸键，荧屏显示操作，进口仪器有的可以升级为荧光定量PCR仪，而国产的目前主要是专用机。而这些仪器附带功能的差别大多都是最近两年新推出来的，价格确是相差几倍。有的进口仪器没有这些附带的特殊功能，从性能和功能上比有的等于或低于国产的。
谢谢！
2、放 PCR 离心管，先把顶盖向上扳，再往前推，露出放样孔，PCR 管
放入前应加好反应体系各组分，再放入 PCR 离心管。
3、反向重复第二步骤将顶盖板向后拉，盖好顶盖。
4、按 F2“Create”新建一个扩增的 PCR 程序。
5、按控制台面右方上下左右方向键，可随意移动编辑位置，输入需要
的温度、时间、循环数等。
6、据 PCR 离心管中加入的反应体积总量，在“Reaction volume”后输入
相应数值，有 Std“1～100ul”和 9600“1~50ul”两档可供选择。
7、按 F1“Start”启动
8、按“INFO”对应键查看 PCR 结束键。
9、扩增完成后按台面左方“Stop”键退出。
10、按“Hist”，可查看上次扩增的历史记录。11、完全退出后，按台面左下方电源开关，拔出插销，切断电源。
PCR技术类似于DNA的天然复制过程，其特异性依赖于与靶序列两端互补的寡核苷酸引物,由变性--退火--延伸三个基本反应步骤构成
