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NEB® Stable Competent E. coli (High Efficiency)

Catalog # Concentration Size List Price Quantity Your Price
C3040H 20 x 0.05 ml $531.00
$477.90
C3040I 6 x 0.2 ml $409.00
$368.10
Catalog # Size List Price Your Price
C3040H 20 x 0.05 ml $531.00
$477.90
C3040I 6 x 0.2 ml $409.00
$368.10
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Chemically competent E. coli cells suitable for high efficiency transformation.

  • Ideal for isolation of plasmid clones containing repeat elements and unstable inserts
  • Useful for isolating and propagating retroviral/lentiviral clones
  • Carries the endA mutation (in contrast to Stbl3) so that isolated plasmids are free of Endonuclease I
  • No dry ice surcharge on competent cell shipments
  • Outgrowth medium included
  • Free of animal products
Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    Tips for Successful Transformations with NEB® Competent Cells
  • Monarch_MiniprepProtocol_VideoThumb
    How to Perform a Transformation with NEB® Competent Cells
  • Monarch_MiniprepProtocol_VideoThumb
    The Mechanism of Transformation with Competent Cells
  • Monarch_MiniprepProtocol_VideoThumb
    Overview of PCR Cloning
  • Monarch_MiniprepProtocol_VideoThumb
    Traditional Cloning Workflow
  • Monarch_MiniprepProtocol_VideoThumb
    NEBUILDER HIFI DNA ASSEMBLY® Removal of 3´ end Mismatches
  • Monarch_MiniprepProtocol_VideoThumb
    Fast transformation plating with glass beads

Chemically competent E. coli cells suitable for high efficiency transformation and isolation of plasmid clones containing repeat elements.

Highlights
  • Recommended host strain for cloning genes into retroviral/ lentiviral vectors 
  • Recommended for cloning of direct repeats and inverted repeat sequences
  • Reduced recombination of cloned DNA (recA1)
  • Efficient transformation of methylated DNA derived from eukaryotic sources or unmethylated DNA derived from PCR, cDNA and many other sources [mcrAΔ(mrr-hsdRMS-mcrBC)]
  • Activity of nonspecific endonuclease I (endA1) eliminated for highest quality plasmid preparations
  • Resistance to phage T1 (fhuA)
  • Suitable for blue/white screening with vectors capable of alpha-complementation
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NEB® 10-beta/Stable Outgrowth Medium B9035SVIAL 4 1 x 25 ml
  pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
  NEB® Stable Competent E. coli (High Efficiency) C3040HVIAL -80 20 x 0.05 ml
  NEB® 10-beta/Stable Outgrowth Medium B9035SVIAL 4 1 x 25 ml
  pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
  NEB® Stable Competent E. coli (High Efficiency) C3040IVIAL -80 6 x 0.2 ml
Features
  • High quality plasmid preparations due to endA mutation
  • T1 phage resistance (fhuA)
  • Value pricing
  • Free of animal products
Application Features
  • Cloning unstable inserts
  • Isolating and propagating retroviral/ lentiviral clones
  • Compatible with NEBuilder HiFi DNA Assembly, Gibson Assembly reactions as well as ligation reactions

NEB stable enables the isolation of plasmid clones containing repetitive DNA elements


Plasmid pUC-5xREP contains five 32-bp repeats, making it unstable in a recombination-proficient strain. A) NEB Stable competent cells or B) Stbl3 competent cells were transformed with 2 µl of a pUC-5xREP Gibson Assembly reaction containing 2.2 ng (0.00125 pmol) pUC19 vector and approximately 1 ng (0.0028 pmol) 5xREP insert. Transformed cultures were plated on LB plates containing 100 µg/ml ampicillin and incubated overnight at 30°C. The next day, colony PCR was performed using M13/pUC polylinker primers to analyze 5xREP insert stability. Figure 1 shows the results of analyzing 33 independent colonies. The correct full-length amplicon is 623bp.
Benefit from the high transformation efficiencies of NEB stable competent E. coli
NEBStable_Stbl3_TE_1217 Transformation efficiencies were compared using manufacturers’ recommended protocols. Values shown are the average of triplicate experiments.
Effect of outgrowth medium on transformation efficiency
C3040_OutgrowthMedium_0517a
50 μl of NEB Stable competent E. coli was transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol with the exception of varying the outgrowth medium. NEB 10-beta/Stable Outgrowth Medium delivers the highest transformation efficiency.

Properties & Usage

Antibiotic for Plasmid Selection

Antibiotics for Plasmid Selection Working Concentration
Ampicillin 100 µg/ml
Carbenicillin 100 µg/ml
Chloramphenicol 33 µg/ml
Kanamycin 30 µg/ml

Shipping Notes

  • Ships on dry ice

Antibiotic Resistance

  • tetracycline
  • streptomycin


Materials Sold Separately
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.
  • Transformation reactions with AmpR plasmids do not require an outgrowth period after addition of Outgrowth Medium. Plasmid selection using antibiotics other than ampicillin requires an outgrowth period of 60 minutes at 30°C before plating on selective media. 30°C or 37°C  may be used for plate incubation, however 30°C is recommended as some constructs may be unstable at elevated temperatures.
Quality Control Assay
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.
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]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
Legal And Disclaimer

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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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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