BL21(DE3) Competent E. coli

Transcription

1 SINGAPORE Home Products BL21(DE3) Competent E. coli BL21(DE3) Competent E. coli NEB does not add a dry ice surcharge to our competent cell shipments. Catalog # Size Concentration C2527I 6 x 0.2 ml/tube C2527H 20 x 0.05 ml/tube Categories: E. coli, E. coli Expression Strains Applications: E.coli Protein Expression, IMPACT Protein Expression, Peptide Ligation Product Information FAQs & Tech Tips Protocols & Manuals Other Tools & Resources Quality & Safety Legal Information Description Properties and Usage Notes Advantages and Features Related Products Description Chemically competent E. coli cells suitable for transformation and protein expression. Highlights Transformation efficiency: 1 5 x 10 7 cfu/µg puc19 DNA T7 Expression Strain Deficient in proteases Lon and OmpT Resistant to phage T1 (fhua2) B Strain Free of animal products Genotype fhua2 [lon] ompt gal (λ DE3) [dcm] hsds λ DE3 = λ sbamhio EcoRI-B int::(laci::placuv5::t7 gene1) i21 nin5 Reagents Supplied The following reagents are supplied with this product: Store at ( C) Concentration puc19 Vector ng/μl SOC Outgrowth Medium 4 1X Advantages and Features Features Routine T7 expression

2 Properties and Usage Antibiotics for Plasmid Selection Ampicillin Carbenicillin Chloramphenicol Kanamycin Streptomycin Tetracycline Working Concentration 100 μg/ml 100 μg/ml 33 μg/ml 30 μg/ml 25 μg/ml 15 μg/ml Storage Temperature -80 C Shipping Notes Ships on dry ice Related Products Companion Products BL21 Competent E. coli Lemo21(DE3) Competent E. coli NiCo21(DE3) Competent E. coli Materials Sold Separately SOC Outgrowth Medium Notes 1. CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling. 2. STORAGE AND HANDLING: Competent cells should be stored at -80 C. Storage at -20 C will result in a significant decrease in transformation efficiency. Cells lose efficiency whenever they are warmed above -80 C, even if they do not thaw. FAQs FAQs 1. Does New England Biolabs offer strains of Competent E.coli suited for protein expression? 2. What are the strain properties (C2527)? 3. Why are there no colonies or no growth in liquid culture (C2527)? 4. Why is induced protein insoluble (C2527)? 5. Why is there no protein visible on gel or no activity (C2527)? 6. What are the solutions/recipes (C2527)? 7. Can I store competent cells at -20 C instead of -80 C? 8. Which kind of transformation tubes should be used? 9. What volume of DNA can be added into competent cells? 10. What is the shelf life for this strain (NEB #C2527H and NEB #C2527I)? 11. Are NEB's competent cells compatible with the Mix & Go" protocol? 12. What is the difference between BL21 and BL21(DE3) competent E.coli cells? 13. How should I store SOC Outgrowth Medium? The SOC I received with my competent cells recommends storage at either room temperature or 4 C, however, when I purchase it as a stand alone product, it recommends storing it at 4 C. Which is better? Protocols Datacards Protocols 1. 5 Minute Transformation Protocol (C2527) 2. Protein Expression Using BL21(DE3) (C2527) 3. Transformation Protocol for BL21(DE3) Competent Cells (C2527)

3 Datacards The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at or fill out the Technical Support Form for appropriate document. C2527Datasheet-Lot21 C2527Datasheet-Lot22 C2527Datasheet-Lot23 C2527Datasheet-Lot24 C2527Datasheet-Lot25 C2527Datasheet-Lot26 C2527Datasheet-Lot27 C2527Datasheet-Lot28 C2527Datasheet-Lot29 C2527Datasheet-Lot30 C2527Datasheet-Lot31 C2527Datasheet-Lot32 C2527Datasheet-Lot33 C2527Datasheet-Lot34 C2527Datasheet-Lot35 C2527Datasheet-Lot36 C2527Datasheet-Lot37 C2527Datasheet-Lot38 C2527Datasheet-Lot42 C2527Datasheet-Lot43 C2527Datasheet-Lot44 C2527Datasheet-Lot C2527Datasheet-Lot C2527Datasheet-Lot C2527Datasheet-Lot C2527Datasheet-Lot C2527Datasheet-Lot C2527IDatasheet-Lot C2527Datasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527IDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot Selection Charts Application Notes Usage Guidelines & Tips Selection Charts E. coli Strains Supplied by NEB Competent E. coli Strains Competent Cell Product Comparison NEB Competent E coli Strains Strain Properties Usage Guidelines & Tips

4 Additional E. coli Strain Genotypes Convenient Formats of Competent Cells Genetic Markers Making Unmethylated (Dam- Dcm-) DNA McrA, McrBC and EcoKI Strain Phenotypes Application Notes Protein Expression with T7 Express Strains C2527 Quality Control Specifications Datacards Certificate of Analysis Safety Data Sheet Quality Control Quality Control Assays The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here. Transformation Efficiency: The competent cells are tested for transformation efficiency and pass minimum release criteria. Transformation efficiency is defined as the number of colony forming units (cfu) which would be produced by transforming 1 μg of plasmid into a given volume of competent cells. Certificate of Analysis The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number] C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_

5 C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ C2527H_I_v1_ Specifications The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version] C2527H_I_v1 Safety Data Sheet The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely. BL21(DE3) Competent E. coli puc19 Vector SOC Outgrowth Medium Datacards The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document. C2527Datasheet-Lot21 C2527Datasheet-Lot22 C2527Datasheet-Lot23 C2527Datasheet-Lot24 C2527Datasheet-Lot25 C2527Datasheet-Lot26 C2527Datasheet-Lot27 C2527Datasheet-Lot28 C2527Datasheet-Lot29 C2527Datasheet-Lot30 C2527Datasheet-Lot31 C2527Datasheet-Lot32 C2527Datasheet-Lot33 C2527Datasheet-Lot34 C2527Datasheet-Lot35 C2527Datasheet-Lot36 C2527Datasheet-Lot37 C2527Datasheet-Lot38 C2527Datasheet-Lot42 C2527Datasheet-Lot43 C2527Datasheet-Lot44 C2527Datasheet-Lot C2527Datasheet-Lot

6 C2527Datasheet-Lot C2527Datasheet-Lot C2527Datasheet-Lot C2527Datasheet-Lot C2527IDatasheet-Lot C2527Datasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527IDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot C2527HDatasheet-Lot Legal and Disclaimers Legal and Disclaimers This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications. For more information about commercial rights, please contact NEB's Global Business Development team at This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.