T7 Express Iq Competent E. coli (High Efficiency)

T7 Express I^q Competent E. coli (High Efficiency)

NEB does not add a dry ice surcharge to our competent cell shipments.
T7 Express lysY/I^qcompetent E.coli (C3013I) is available as a superior alternative to T7 Express I^q competent E.coli (C3016 ). The additional lysY gene expresses a phage T7 lysozyme variant that inhibits basal T7 RNA polymerase activity. The net result of employing lysY/I^q cells as a host strain is greater clone stability and equivalent or improved expression of recombinant protein.

Catalog # C3016 was discontinued on March 31, 2015 ;

Product Information
Enhanced BL21 E. Coli derivative, chemically competent E. coli cells suitable for high efficiency transformation and protein expression.
Highlights
- Transformation efficiency: 0.6-1 x 10⁹cfu/μg pUC19 DNA
- Enhanced BL21 derivative for T7 expression
- T7 RNA Polymerase in the lacoperon - no lambda prophage
- Tight control of expression by lacl^q allows potentially toxic genes to be cloned
- Deficient in proteases Lon and OmpT
- Resistant to phage T1 (fhuA2), Cam, Nit
- Does not restrict methylated DNA (McrA^-, McrBC^-, EcoBr^-, Mrr^-)
- B Strain
- Free of animal products
- lac^q on miniF for stability
- no Cam requirement
Genotype
MiniF lacI^q(Cam^R) / fhuA2 lacZ::T7 gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--Tet^S)2 [dcm] R(zgb-210::Tn10--Tet^S) endA1 Δ(mcrC-mrr)114::IS10
Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration

SOC Outgrowth Medium 4 1 X

pUC19 Vector -20 0.05 ng/µl

Product Categories:

Discontinued Products

Applications:

Peptide Ligation
- Advantages and Features
  Features
  
  T7 expression
  
  Protease deficient
  
  Reduced basal expression
  
  Application Features
  
  Transformation of a toxic mammalian clone into E. coli hosts. A T7 expression plasmid and the same plasmid containing a gene encoding a toxic mammalian protein were transformed into each host. Comparison of the relative transformation efficiencies demonstrates that the T7 Express hosts provide the levels of control necessary for transformation of potentially toxic clones. BL21(DE3) could not be transformed with the toxic clone.
  
  T7-controlled expression of a non-toxic protein in E. coli hosts. A T7 expression plasmid containing a gene encoding an E. coli protein was transformed into each host, grown to 0.6 OD and induced for 3 hours. Comparison of soluble extracts from uninduced (-) and induced (+) cells shows superior control of basal expression in the T7 Express hosts while maintaining high levels of induced expression.
  
  * Ideally, DNA for transformation should be purified and resuspended in water or TE. However, up to 10 µl of DNA directly from a ligation mix can be used with only a two-fold loss of transformation efficiency. Where it is necessary to maximize the number of transformants (e.g. a library), a purification step, either a spin column or phenol/chloroform extraction and ethanol precipitation should be added.
  
  Effect of heat shock time on T7 Express I^q competent E.coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except heat shock time varied from 0 to 80 seconds.
  
  Effect of DNA incubation time on T7 Express I^q competent E.coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except DNA incubation time varied from 0 to 40 minutes.
- Properties & Usage
  Antibiotics for Plasmid Selection Working Concentration
  
  Ampicillin 100 μg/ml
  
  Carbenicillin 100 μg/ml
  
  Kanamycin 30 μg/ml
  
  Tetracycline 15 μg/ml
  
  Streptomycin 25 μg/ml
  
  Storage Temperature
  -80°C
  Shipping Notes
  
  Ships on dry ice
  
  Antibiotic Resistance
  
  cam
  
  nit
- Related Products
  Companion Products
  
  T7 Express Competent E. coli (High Efficiency)
  
  T7 Express lysY Competent E. coli (High Efficiency)
  
  T7 Express lysY/I^q Competent E. coli (High Efficiency)
  
  Materials Sold Separately
  
  SOC Outgrowth Medium
- Product Notes
  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.
  
  CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.
  
  Maintenance of the miniF plasmid does not require antibiotic selection. If chloramphenicol is added, use 10 µg/ml final concentration.
Protocols, Manuals & Usage
- Protocols
  High Efficiency Transformation Protocol (C3016)
  
  5 Minute Transformation Protocol (C3016)
  
  Protocol for Expression Using T7 Express Iq (C3016)
- Usage Guidelines
  Additional E. coli Strain Genotypes
  
  Convenient Formats of Competent Cells
  
  Genetic Markers
  
  McrA, McrBC and EcoKI Strain Phenotypes
- Application Notes
  Protein Expression with T7 Express Strains C3016
- 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 [email protected] or fill out the Technical Support Form for appropriate document.)
  
  C3016Datasheet-Lot6
Tools & Resources
- Selection Charts
  Characteristics of Select E.coli Strains
  
  Competent Cell Selection Guide
  
  Strain Properties
FAQs & Troubleshooting
- FAQs
  Why are there no colonies or no growth in liquid culture (C3016)?
  
  Why is there no protein visible on gel or no activity (C3016)?
  
  Why is induced protein insoluble (C3016)?
  
  What are the solutions/recipes (C3016)?
  
  What are the strain properties (C3016)?
  
  Can I store competent cells at -20°C instead of -80°C?
  
  What is the difference between NEB #C3016H and NEB #C3016I?
  
  What is the optimal heat shock time for this strain (NEB #C3016H and NEB #C3016I)?
  
  How long should I incubate cells on ice after DNA has been added (NEB #C3010H and NEB #C3010I)?
  
  Is T7 Express I^q (NEB #C3016H and NEB #C3016I) compatible with auto-induction procedures?
  
  Which kind of transformation tubes should be used?
  
  What volume of DNA can be added into competent cells?
  
  What is the shelf life for this strain (NEB #C3016H and NEB #C3016I)?
  
  Are NEB's competent cells compatible with the “Mix & Go" protocol?
Quality, Safety & Legal
- Quality Control Assays
  
  Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  T7 Express I^q Competent E. coli (High Efficiency)
  
  SOC Outgrowth Medium
  
  pUC19 Vector
- 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 [email protected] or fill out the Technical Support Form for appropriate document.)
  
  C3016Datasheet-Lot6
- 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 [email protected].
  
  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.