Académique Documents
Professionnel Documents
Culture Documents
NEB 5-alpha F´ I q
1–3 x 10 9
K12 l l l – – l – l l – – tet
NEB Express I q
0.6–1 x 10 9
B l – l – – l l – – – – cam, nit
T7 Express 0.6–1 x 10 9
B l – – – – l l – – l – nit
T7 Express I q
0.6–1 x 10 9
B l – l – – l l – – l – cam, nit
T7 Express lysY/I q
0.6–1 x 10 9
B l – l l – l l – – l – cam, nit
SHuffle ™
1 x 10 6
K12 l – l – – – – l – – l str, spec
SHuffle T7 ™
1 x 10 6
K12 l – l – – – – l – l l str, spec
cam, str,
SHuffle™ T7 lysY 1 x 106 K12 l – l l – – – – – l l
spec
SHuffle™ Express 1 x 107 B l – l – – l l – – – l spec
SHuffle™
1 x 107 B l – l – – l l – – l l spec(6)
T7 Express
SHuffle™ T7
1 x 107 B l – l l – l l – – l l cam, spec(6)
Express lysY
BL21(DE3) 1–5 x 107 B l – – – – – l – – l – –
Lemo21(DE3) 1–3 x 10 7
B l – – l – – l – – l – cam
(1) TE are given high-quality for high efficiency chemically competent strains. TE for electrocompetent strains are 1-4 x 1010 cfu/µg. TE for subcloning strains are >1 x 106 cfu/µg.
(2) Important for high-quality plasmid preparation.
(3) Lacks Lon and OmpT protease activity.
(4) Constitutively expresses a chromosomal copy of the disulfide bond isomerase DsbC.
(5) nit = nitrofurantoin, tet = tetracycline, cam = chloramphenicol, str = streptomycin, spec = spectinomycin
(6) Resistance to low levels of streptomycin may be observed.
2
Convenient Formats Visit the technical reference section
of www.neb.com for more
information, including tips for
NEB competent cell strains are available in various formats for your convenience. enhancing transformation efficiencies.
All are available as 50 µl single-use transformation tubes and many are offered in larger,
200 µl tubes for multiple simultaneous reactions. Our most popular cloning strains are
available as electrocompetent cells. SOC outgrowth media and control plasmids are
provided with many of these formats. For even greater value, NEB 5-alpha is also avail-
able in a lower efficiency, subcloning format.
Cloning Formats
NEB 5-alpha l l l l l
NEB 5-alpha F´ I q l l l
NEB 10-beta l l l l
– –
dam /dcm l l l
NEB Express I q l l l
T7 Express l l l
T7 Express I q l l l
T7 Express lysY l l l
T7 Express lysY/I q l l l
T7 Express Crystal l l l
SHuffle™ l
SHuffle™ T7 l
SHuffle™ T7 lysY l
SHuffle™ Express l
SHuffle™ l
T7 Express
SHuffle™ T7 l
Express lysY
BL21(DE3) l l
Lemo21(DE3) l
www.neb.com 3
CLONING & PROTEIN EXPRESSION
Cloning Strains
NEB’s growing line of competent cells includes several popular strains for cloning and Advantages:
protein expression, in addition to strains with unique properties including fast colony
• Specialized cloning applications
growth, tight control of expression and disulfide bond formation. Our cloning strains
include derivatives of the industry standards, DH5a™ and DH10B™. NEB Turbo is • Strains for cloning toxic genes
unique to NEB and allows colony growth after 6.5 hours. NEB’s dam–/dcm– strain • Free of animal products
enables Dam and Dcm methylation free plasmid growth. Our cells are all extensively • T1 phage resistance (fhuA2)
tested for phage resistance, antibiotic resistance and sensitivity, blue/white screening and
• Media and control plasmid included
transformation efficiency. High efficiency, 5 Minute and Electroporation Protocols
are provided when applicable. • A variety of convenient formats
• Bulk sales available for
Cloning custom packaging
CHARACTERISTICS STRAIN
With NEB Turbo, colonies are visible after only 8 hours. Ligation products were transformed into 50 µl of NEB Turbo Competent E. coli and
plated on LB/Amp. Plates were incubated for 8 hours, 10 hours and 12 hours at 37°C. NEB Turbo features fast colony growth and blue/white
selection to simplify cloning experiments.
4
Protein Expression Strains
NEB also offers a wide variety of competent cell strains ideal for many protein expres- Advantages:
sion applications. These strains address the needs of protein expression control, toxic
• Deficient in proteases Lon and OmpT
protein expression, cytoplasmic disulfide bond formation, difficult targets and crystal-
lography. NEB Express, T7 Express and SHuffle™ strains are available with varying • Do not restrict methylated DNA
levels of control. I q strains feature the added control of IPTG-induced expression of • Free of animal products
non-T7 plasmids by lacI q . Only NEB offers the exceptional control of expression from
• T1 phage resistance ( fhuA2 )
the lysY gene that reduces basal expression from T7 strains without inhibiting IPTG-
induced expression. Our Lemo21(DE3) strain features tunable T7 expression for • Media and control plasmid included
difficult targets such as membrane proteins and proteins prone to insoluble expression. • A variety of convenient formats
Each strain is provided with a detailed protocol for optimal expression.
• Bulk sales available for
custom packaging
Protein Expression
CHARACTERISTICS STRAIN
• Tunable T7 Expression
• Expression of difficult targets including membrane proteins, Lemo21(DE3)
toxic proteins and proteins prone to insoluble expression
• Control of IPTG induced expression from Plac , Ptac and Ptrc NEB Express I q Competent E. coli
• T7 expression
T7 Express I q Competent E. coli
• Reduced basal expression
• T7 expression
T7 Express lysY Competent E. coli
• Better reduction of basal expression
• T7 expression
• Lowest basal expression
T7 Express lysY/I q Competent E. coli Not sure which strain
• T7 expression
T7 Express Crystal Competent E. coli
to use? – Try one of
• SeMet labeling of protein for crystallography
www.neb.com 5
ADDITIONAL
CLONING CONVENIENCE
& PROTEIN EXPRESSION
8,000 5,000
Improved transformation of toxic clones with
T7 Expression strains from NEB
4,000
Relative Activity
6,000
PhoA Activity (units)
60
3,000
Percent Transformed (%)
40 4,000
2,000
2,000
20
1,000
0
0 q q Origami (DE3) SHuffle T7 SHuffle T7 0
BL21 (DE3) T7 Express lysY lysY/l l 0 100 500 1,000
Express
Transformed Strain µM rhamnose
A T7 expression plasmid and the same plasmid containing a gene Plasmodium falciparum chitinase (PfCHT1) with three cysteines was Lemo21(DE3) System™ enables simple, rapid optimization of mem-
encoding a toxic mammalian protein were transformed into each expressed from a plasmid under the regulation of a T7 promoter. After brane protein expression.
host. Comparison of the relative transformation efficiencies demon- induction, cells were harvested and crude cell lysates were prepared.
strates that the T7 Express hosts provide the levels of control PfCHT1 was assayed using a chromogenic substrate (CalBioChem
necessary for transformation of potentially toxic clones. BL21(DE3) #474550) and standardized to protein concentration using Bradford
could not be transformed with the toxic clone. reagent.
6
ORDERING INFORMATION
Cloning Strains
PRODUCT NEB # SIZE
NEB Turbo Competent E. coli C2984H 20 x 0.05 ml
NEB Turbo Competent E. coli C2984I 6 x 0.2 ml
NEB Turbo Electrocompetent E. coli C2986K 6 x 0.1 ml
NEB 10-beta Competent E. coli C3019H 20 x 0.05 ml
NEB 10-beta Competent E. coli C3019I 6 x 0.2 ml
NEB 10-beta Electrocompetent E. coli C3020K 6 x 0.1 ml
NEB 5-alpha Competent E. coli C2987H 20 x 0.05 ml
NEB 5-alpha Competent E. coli C2987I 6 x 0.2 ml
NEB 5-alpha Competent E. coli (Subcloning Efficiency) C2988J 6 x 0.4 ml
NEB 5-alpha Electrocompetent E. coli C2989K 6 x 0.1 ml
NEB 5-alpha F´ I q Competent E. coli C2992H 20 x 0.05 ml
NEB 5-alpha F´ I Competent E. coli
q
C2992I 6 x 0.2 ml
dam – /dcm – Competent E. coli C2925H 20 x 0.05 ml
dam – /dcm – Competent E. coli C2925I 6 x 0.2 ml
Component Sold Separately: SOC Outgrowth Medium B9020S 4 x 25 ml
info@ca.neb.com BOSTON, MA
PERMIT NO. 54302
China, People’s Republic New England Biolabs, Inc.
New England Biolabs (Beijing), Ltd. 240 County Road
Telephone: 010-82378265/82378266 Ipswich, MA 01938-2723
info@neb-china.com
Germany
New England Biolabs GmbH
Free Call: 0800/246 5227
info@de.neb.com
United Kingdom
New England Biolabs (UK), Ltd.
Call Free 0800 318486
info@uk.neb.com
Japan
New England Biolabs Japan, Inc.
Telephone: +81 (0)3 5669 6191
info@neb-japan.com
www.neb.com
CEL
Version 2.0