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- http://lucigen.com/store/docs/manuals/MA031-OverExpress-Chemicallycompetent-Cells.pdf
- 60435-1
- 2026年01月11日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-80º C
- 英文名：
HI-Control® BL21(DE3) Chemically Competent Cells
- 库存：
大量
- 供应商：
中北林格
- 规格：
12 rxns (SOLOs)
HI-Control™ BL21(DE3) and HI-Control™ 10G Competent Cells
产品说明书
- Induce high level protein expression from lac based promoters (T7, T5-lac, tac, trc, lac)
- Tightly regulate expression from T7 promoters with HI-Control BL21(DE3) Chemically Competent Cells
- Clone targets and reduce background protein expression with HI-Control 10G Chemically Competent Cells
The HI-Control™ strains are based on E. cloni10G ( DH10B) and BL21(DE3) strains. These strains harbor a plasmid bearing a constitutivelacI repressor gene, which provides tight control over promoters containing thelacO operator.
Reduce leaky expression: The LacI repressor protein binds to the lacO operator, blocking transcription of downstream genes. In the absence of an induction agent such as lactose sugar or IPTG, target protein expression from the adjacent promoter is greatly reduced.
Obtain high protein yields: Both 10G and BL21(DE3)-based strains express high levels of protein. For expression from a T7 promoter, select the HI-Control BL21(DE3) strain. Other lac-based promoters like T5-lac, tac, trc, and lac, can be used in either HI-Control strain.
Achieve high efficiency transformations: Highly efficient competent cells give excellent results from small amounts of plasmid DNA.
| Tightly Control Protein Expression | ||||
| T7 Promoters | Non-T7 lac promoters (T5-lac, tac, trc, lac) |
Clone with low background target expression | Transformation efficiencies | |
| HI-Control BL21(DE3) |
≥ 1×107 cfu/µg | |||
| HI-Control 10G |
≥ 1×109 cfu/µg | |||
Genotypes:
HI-Control™ BL21 (DE3): F- ompT hsdSB (rB- mB-) gal dcm (DE3) Mini-F lacIq1(GentR)
HI-Control™ 10G: mcrA Δ(mrr-hsdRMS-mcrBC) endA1 recA1 ?80dlacZΔM15 ΔlacX74 araD139 Δ (ara,leu)7697 galU galK rpsL (StrR) nupG λ− tonA Mini-F lacIq1 (GentR)
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Ordering
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Specifications
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Manuals
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Resources
Competent Cells include Control DNA and Recovery Medium, and are packaged as SOLOs (1 transformation per tube). Recovery Medium is also available separately. The specified transformation efficiencies are with pUC DNA, unless indicated otherwise.
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文献和实验TOP10 chemically competent cells
Overview This protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the Bloom05 patent as well. This protocol has been tested on TOP10, MachI and BL21(DE3) cells
Transforming chemically competent cells
Method Thaw TSS cells on ice. Add DNA, pipette gently to mix (1μl of prepped plasmid is more than enough). Note: If you are adding small volumes (~1μl), be careful to mix the culture well. Diluting the plasmid
Preparing chemically competent cells
Materials Plate of cells to be made competent TSS buffer LB media Ice Glassware & Equipment Falcon tubes 500μl Eppendorf tubes, on ice 200ml conical flask 200μl pipetman or repeating
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