Now supplied with outgrowth medium optimized for NEB 10-beta & NEB Stable Competent E.coli ; please do not use SOC outgrowth medium.
NEB 10-beta Competent E. coli is a derivative of the popular DH10B. It is T1 phage resistant and endonuclease I (endA1) deficient for high- quality plasmid preparations.
High efficiency strain ideal for cloning large plasmids and BACs
Chemically competent E. coli cells suitable for high efficiency transformation in a wide variety of applications.
Highlights
Transformation efficiency: 1–3 x 109 cfu/µg pUC19 DNA
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 without IPTG by a-complementation of the b-galactosidase gene
Effect of Plasmid Size on Transformation Efficiency:
NEB 10-beta chemically competent cells are more efficiently transformed with large plasmids than NEB 5-alpha cells. The difference in TE between the two cell lines increases with the size of the plasmid being transformed.
Effect of heat shock time on NEB 10-beta 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 NEB 10-beta 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.
Effect of outgrowth medium on transformation efficiency:
50 μl of NEB 10-beta 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.Benefit from the high transformation efficiencies of NEB 10-beta:
Transformation efficiencies were compared using manufacturers’ recommended protocols. Values shown are the average of triplicate experiments.
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.
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.)
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.
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]
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]
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.)
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Trademarks
ONESHOT® and MAX EFFICIENCY® are registered trademarks of Thermo Fisher Scientific. DH10B™ is a trademarks of Thermo Fisher Scientific.
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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.
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