NEB® 5-alpha Competent E. coli (High Efficiency)

Catalog #	Concentration	Size
C2987I	Not Applicable	6 x 0.2 ml/tube
C2987P	Not Applicable	1 x 96 well plate (20 µl/well)
C2987H	Not Applicable	20 x 0.05 ml/tube
C2987R	Not Applicable	1 x 384 well plate (10 µl/well)
C2987U	Not Applicable	96 x 50 µl/tube (12 x 8-tube strips)
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Product Information

Chemically competent E. coli cells suitable for high efficiency transformation in a wide variety of applications. Available in 5 convenient formats:

Single-use 50 μl vials
200 μl vials
96-well plate: packed in a 96-well divisible polypropylene plate (Eppendorf twin.tec^® PCR divisible plates)
384-well plate
12 x 8-tube strips: packed in 12 x 8-Strip Microplate Cluster Tube (Costar™ Cluster Tube System)

For C2987H, C2987I, C2987P and C2987U, SOC Outgrowth Medium is provided.
For C2987R, Outgrowth Medium 1.5 is provided.

Highlights

DH5α™ derivative
Free of animal products
Transformation efficiency:
For C2987H, C2987I, C2987P and C2987U: 1 - 3 x 10⁹ cfu/μg pUC19 DNA
For C2987R: 1 - 5 x 10⁸ cfu/μg pUC19 DNA
Efficient transformation of unmethylated DNA derived from PCR, cDNA and many other sources (hsdR)
Activity of nonspecific endonuclease I (endA1) eliminated for highest quality plasmid preparations
Resistance to phage T1 (fhuA2)
Suitable for blue/white screening by α-complementation of the β-galactosidase gene
Reduced recombination of cloned DNA (recA1)
K12 Strain

Genotype

fhuA2 Δ(argF-lacZ)U169 phoA glnV44 Φ80 Δ(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17

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:: Cloning Competent Cell Strains Products

Applications:: USER^® Cloning,; Transformation,; Applications of USER® and Thermolabile USER II Enzymes

Advantages and Features
Features
- Versatile cloning strain
- DH5α™ derivative
Application Features

DNA Effects on Transformation Efficiency and Colony Output:
The optimal amount of DNA to use in a transformation reaction is lower than commonly recognized. Using clean, supercoiled pUC19, the efficiency of transformation is highest in the 100 pg-1 ng range. However, the total colonies which can be obtained from a single transformation reaction increase up to about 100 ng.
Effect of heat shock time on NEB 5-alpha 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 5-alpha 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 5-alpha 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 SOC outgrowth medium delivers the highest transformation efficiency.
Benefit from the high transformation efficiencies of NEB 5-alpha:
The transformation efficiencies of NEB 5-alpha and DH5α were compared using each manufacturers' recommended protocols. Values shown are the average of triplicate experiments.

Properties & Usage

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

none

Related Products
Companion Products
Materials Sold Separately
- SOC Outgrowth Medium
Product Notes
1. For C2987H, C2987I, C2987P and C2987U, SOC Outgrowth Medium is provided.
  For C2987R, Outgrowth Medium 1.5 is provided.
2. CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.
3. 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.
  
  SOC medium can be stored at either 4°C or Room Temperature depending on how fast it will be used. Storing at Room Temperature is convenient and adequate for short term usage (weeks to a couple of months). For long term storage, we recommend storing at 4°C. Please note that Outgrowth Medium 1.5 supplied with C2987R (1 x 384 well plate format) must only be stored at Room Temperature or crystals will form.