Xtreme Efficiency DH10ß

Electrocompetent Cells, most

commonly used E.coli strains for high efficiency
plasmid transformation by electroporation.

X treme Efficiency
Electrocompetent cells are the
DH10ß

most commonly used E.coli strains for

Box 2 | Storage

high efficiency plasmid transformation Store at -80°C

by electroporation. Xtreme Efficiency
cells are prepared with our finely tuned It has been observed that long term storage of competent cells at -80°C
procedures and are quality controlled by can actually cause the cells to adapt by increasing the recombination rate.
direct comparison to other leading brands Therefore, Allele prepares its competent cells frequently at small scales. It
of similar products. Xtreme Efficiency is recommended that competent cells be used within 3 months
DH10ß is suitable for propagation of after arrival.
plasmids of all sizes and preparations,
especially for large vectors or inserts.
Allele’s competent cells are quality
controlled by direct comparison to other leading brands of
Protocol
similar products in transformation of ligated plasmids. These
Note: Use ligated or plasmid DNA free of phenol, salts,protein,
competent cells are extremely efficient, consistently yielding
and detergents to obtain maximum transformation efficiency.
above 5 x 10 cfu/μg of supercoiled pUC 19 plasmid DNA.
9

They are suitable for large plasmid constructs and various For ligated reactions, precipitate the DNA with ethanol in the
electro cuvettes. To avoid repeated freezing in dry ice and presence of 5 - 10 μg tRNA and resuspend in TE Buffer (10
alcohol bath competent cells are conveniently packaged in mM Tris HCl, pH 7.5, 1 mM EDTA)
60 μl aliquot. Colonies can be analyzed using blue/white
screening when grown in the presence of X-gal. 1) Add 1 - 3 μL of the DNA to a microcentrifuge tube
on ice.
2) Thaw Xtreme Efficiency DH10B on ice.
3) When cells are thawed, mix cells by tapping gently.
Box 1 | Product List Add 25 μL of cells to each chilled microcentrifuge tube
containing the DNA.
Xtreme Efficiency DH10ß
Electrocompetent Cells 4) Pipette the cell / DNA mixture into a chilled 0.1 cm
ABP-CE-XDHE005 5 x 60μl
cuvette. Gently tap the cuvette to ensure that the mixture
makes contact all the way across the bottom of the cuvette
ABP-CE-XDHE025 25 x 60μl
chamber.Avoid formation of bubbles.
ABP-CE-XDHE050 50 x 60μl
5) Electroporate the sample at 1.6 kV, 200 Ω 25 μF.
6) Add 1 ml of S.O.C. medium to the cells in the
cuvette, and transfer the solution to a 15 ml culture tube.
Genotype 7) Shake at 225 rpm (37°C) for 1 hour.
F-mcrA .(mrr-hsdRMS-mcrBC) φ80lacZ.M15 .lacX74
recA1 endA1 araD139 .(ara, leu)7697 galU galK λ- rpsL
nupG

Related Products
 Allele Competent Cells Spreading Beads
 Provide convenience and to increase
efficiency when spreading transformed bacterial
or yeast cells.
 SC33 High Efficiency Competent Cells
 Ideal for generation of high-difficulty constructs
and propagation of large-size or low concentration
plasmids.

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