Home Competent Cells NEB^® Stable Competent E. coli (High Efficiency)

NEB^® Stable Competent E. coli (High Efficiency)

Supplied with outgrowth medium optimized for NEB 10-beta & NEB Stable Competent E.coli; please do not use SOC outgrowth medium.

Now available in 96-well plate format.

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

Ideal for isolation of plasmid clones containing repeat elements and unstable inserts
Available in single-use vials, 200 μl vials and 96-well plate
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

Catalog #	Cat #	Concentration	Conc	Size	List Price
C3040H	Not Applicable	20 x 0.05 ml	$400.00	$400.00	Sign In Sign In
C3040I	Not Applicable	6 x 0.2 ml	$308.00	$308.00	Sign In Sign In
C3040P	Not Applicable	1 96-well plate (20 μl/well)	$700.00	$700.00	Sign In Sign In

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Product Information

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

Transformation Efficiency

For C3040I and C3040H: 1-3 x 10⁹ cfu/µg pUC19 DNA
For C3040P: 1–3 x 10⁸cfu/µg pUC19 DNA

Genotype

F' proA⁺B⁺ lacI^q ∆(lacZ)M15 zzf::Tn10 (Tet^R)/∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15 e14- Φ80dlacZ∆M15 recA1 relA1 endA1 nupG rpsL (Str^R) rph spoT1 ∆(mrr-hsdRMS-mcrBC)

This product is related to the following categories:: Cloning Competent Cell Strains

This product can be used in the following applications:: High-throughput cloning and automation solutions,; Transformation

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

C3040H

-80

NEB^® 10-beta/Stable Outgrowth Medium	B9035SVIAL	4	1 x 25 ml	Not Applicable
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	Not Applicable

C3040I

-80

NEB^® 10-beta/Stable Outgrowth Medium	B9035SVIAL	4	1 x 25 ml	Not Applicable
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	Not Applicable

C3040P

-80

pUC19 Vector	N3041AVIAL	-20	1 x 0.025 ml	50 pg/µl
NEB^® 10-beta/Stable Outgrowth Medium	B9035SVIAL	4	1 x 25 ml	Not Applicable
NEB^® Stable Competent E. coli (High Efficiency)	C3040PVIAL	-80	1 x 96-well plate (20 μl/well)	Not Applicable

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

Advantages and Features
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
Figure 1: 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. This figure shows the results of analyzing 33 independent colonies. The correct full-length amplicon is 623bp.

Figure 2: Benefit from the high transformation efficiencies of NEB stable competent E. coli

Transformation efficiencies were compared using manufacturers’ recommended protocols. Values shown are the average of triplicate experiments.

Figure 3: Effect of outgrowth medium on transformation efficiency

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.
Related Products
Materials Sold Separately
- NEB® 10-beta/Stable Outgrowth Medium
Product Notes
1. 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.
2. CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.
3. Transformation reactions with Amp^R 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.