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MultiShot™ StripWell TOP10 Chemically Competent E. coli (Invitrogen™)

MultiShot StripWell TOP10 chemically competent E. coli cells are cloning competent cells that packaged in a rack containing 12 strips of 8 tubes to increase productivity for medium-throughput bacterial transformations. TOP10 competent E. coli cells are ideal for high-efficiency cloning and plasmid propagation. They allow stable replication of high-copy number plasmids and are the same competent cells that come with many of our cloning kits.

Versatile TOP10 cloning capabilities
MultiShot FlexPlate TOP10 competent E. coli cells are the laboratory workhorse for chemical transformations as they are compatible with a broad spectrum of DNA types for transformation. The TOP10 strain is useful for many cloning applications and provides transformation efficiencies in the MultiShot FlexPlate format of >1 x 108 cfu/µg with control plasmid DNA.

Key features of the MultiShot FlexPlate TOP10 Competent Cells include:
hsdR for efficient transformation of unmethylated DNA from PCR amplifications
mcrA for efficient transformation of methylated DNA from genomic preparations
lacZΔM15 for blue/white color screening of recombinant clones
endA1 for cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by endonuclease I
recA1 for reduced occurrence of non-specific recombination in cloned DNA

MultiShot StripWell format—flexible for no waste
Each StripWell of eight connected tubes is designed with the same single-tube, single-use features as our One Shot format tubes. This format allows for all steps of the transformation protocol, up to plating, to take place in the same tube, thereby saving time and preventing contamination. After addition of DNA, MultiShot StripWell competent cells can be transformed by heat-shock at 42°C using a water bath, all in the same tube.

The StripWell tubes can be cut to permit transformations in a single tube, eliminating wasted aliquots of cells and avoiding freeze-thaws that can result in reduced transformation efficiency.

Key features of the MultiShot StripWell format include:
Increased productivity—for medium throughput experimentation
Maximal flexibility—transform as few as 1 tube and as many as 96 from the StripWell rack
Familiar convenience—no aliquotting, add DNA directly to cells, heat shock, and recover in the same tube
Peace of mind—single-use transformation minimizes contamination

Genotype: F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara, leu)7697 galU galK rpsL (StrR) endA1 nupG

Find the strain and format that you need
We offer many other E. coli strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs. If you require other high-throughput transformation options, choose from our collection of MultiShot formatted comp cells or email us for custom formatting options.

TOP10 Electrocomp™ Kit (Invitrogen™)

TOP10 E. coli are provided at a transformation efficiency of 1 × 1010 cfu/µg supercoiled DNA and are ideal for high-efficiency cloning and plasmid propagation. They allow stable replication of high-copy number plasmids.

The genotype of TOP10 cells is similar to the DH10B™strain, and offers the following features:

hsdR for efficient transformation of unmethylated DNA from PCR amplifications
mcrA for efficient transformation of methylated DNA from genomic preparations
lacZΔM15 for blue/white color screening of recombinant clones
endA1 for cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by Endonuclease I
recA1 for reduced occurrence of non-specific recombination in cloned DNA

One Shot® TOP10 Kits are available with either chemically competent or electrocompetent E. coli to fit your specific transformation needs. TOP10 E. coli are also available in the high-throughput MultiShot™ format.

Note: This kit contains enough electrocompetent cells for 25 x 40 µL rxns or 50 x 20 µL rxns. Please see the product manual for reaction scale recommendations.

Genotype:
F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL endA1 nupG

MultiShot™ FlexPlate Mach1™ T1R Competent Cells (Invitrogen™)

MultiShot FlexPlate Mach1 T1R chemically competent E. coli cells are cloning competent cells that are pre-aliquoted in a 96-well PCR plate to increase transformation productivity. The FlexPlate format can be divided down to any combination of 8-well segments for flexibility in transformation throughput. MultiShot FlexPlate Mach1 T1R competent E. coli cells are exceptionally fast-growing chemically competent cells, accelerating the time to clone evaluation.

All of our T1R strains have the tonA (fhuA) deletion in their genotype that confers resistance to T1 and T5 phage. T1 bacteriophage spread rapidly and lyse E. coli hosts, which are commonly used for cloning and library construction. A resistant genotype offers added security to protect valuable clones and libraries. This is especially important for genome and sequencing centers where T1 phage infection would be catastrophic.

Speedy Mach1 T1R cloning capabilities
MultiShot FlexPlate Mach1 T1R competent E. coli cells are fast growing. Mach1 colonies are clearly visible within eight hours of plating the transformation mix, enabling you to plate and pick colonies the same day. Perform mini-preps after only four hours of growth from an overnight colony—saving an entire day in your experiments. The Mach1 T1R strain is useful for many cloning applications and provides transformation efficiencies of >1 x 108 cfu/µg with control plasmid DNA.

Key features of the MultiShot FlexPlate Mach1 T1R Competent Cells include:
endA1 for cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by endonuclease I
recA1398 for reduced occurrence of non-specific recombination in cloned DNA
hsdR for efficient transformation of unmethylated DNA from PCR amplifications
tonA (fhuA) for T1 and T5 phage resistance
lacZΔM15 for blue/white color screening of recombinant clones

MultiShot FlexWell format—increase transformation productivity
The FlexPlate is a 96-well PCR plate that can be used in automated liquid handling systems or divided into any combination of 8-well segments for flexibility in transformation throughput. Addition of DNA to the MultiShot FlexPlate Competent Cells can be done quickly with a multichannel pipet or a liquid handling system. After addition of DNA, MultiShot FlexPlate Competent Cells can efficiently be transformed by heat-shock using a water bath, heat block, or thermal cycler.

Key features of the MultiShot FlexPlate format include:
Increase throughput—compatible with automated liquid handling platforms
Flexible—transform in increments of 8 wells to entire 96-well plate
Convenient—competent cells pre-aliquoted into 96-well plate, ready for transformation

Genotype: F- φ80(lacZ)ΔM15 ΔlacX74 hsdR(rK-mK+) ΔrecA1398 endA1 tonA

Find the strain and format that you need
We offer many other E. coli strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs. If you require other high-throughput transformation options, choose from our collection of MultiShot formatted comp cells or email us for custom formatting options.

ElectroMAX™ DH10B Cells (Invitrogen™)

ElectroMAX DH10B Cells are electrocompetent E. coli cells offering the highest transformation efficiencies of >1 x 1010 cfu/µg plasmid DNA. ElectroMAX DH10B Cells are ideal for applications requiring high transformation efficiencies, such as with cDNA or gDNA library construction. ElectroMAX DH10B Cells provide:

• High-efficiency transformation to maximize clones
• Enhanced genetic markers for cDNA or gDNA cloning capabilities

Highest transformation efficiencies
ElectroMAX DH10B Cells offer the highest transformation efficiencies available with >1 x 1010 transformants/µg of control DNA in a 20 µL reaction. These high transformation efficiencies make ElectroMAX DH10B Cells ideal for efficient cloning of both prokaryotic and eukaryotic genomic DNA and efficient plasmid rescue from eukaryotic genomes. ElectroMAX DH10B Cells are also suitable for construction of gene banks, for generation of cDNA libraries using plasmid-derived vectors, and for situations when there is limited amount of DNA.

Note: A high-voltage electroporation apparatus capable of generating field strengths of 16 kV/cm is required.

Versatile cloning capabilities
ElectroMAX DH10B Cells have the following features:

• Φ80lacZΔM15 marker provides α-complementation of the β-galactosidase gene allowing blue/white screening on agar plates containing X-gal or Bluo-gal
mcrA genotypic marker and the mcrBC, mrr deletion allows for cloning DNA that contains methylcytosine and methyladenine
endA1 mutation enables increased plasmid yield and quantity

Genotype: F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139Δ(ara, leu)7697 galU galK λ-rpsL nupG

Find the strain and format that you need
DH10B cells are available in both electrocompetent and chemically competent formats. Our ElectroMax DH10B T1R Cells and MegaX DH10B T1R Electrocomp Cells offer the benefit of T1 and T5 phage resistance.

MultiShot™ StripWell Mach1™ T1 Phage-Resistant Chemically Competent E. coli (Invitrogen™)

MultiShot StripWell Mach1 T1R chemically competent E. coli cells are cloning competent cells that are packaged in a rack containing 12 strips of 8 tubes to increase productivity for medium-throughput bacterial transformations. MultiShot StripWell Mach1 T1R competent E. coli cells are exceptionally fast-growing chemically competent cells, accelerating the time to clone evaluation.

All of our T1R strains have the tonA (fhuA) deletion in their genotype that confers resistance to T1 and T5 phage. T1 bacteriophage spread rapidly and lyse E. coli hosts, which are commonly used for cloning and library construction. A resistant genotype offers added security to protect valuable clones and libraries. This is especially important for genome and sequencing centers where T1 phage infection would be catastrophic.

Speedy Mach1 T1R cloning capabilities
MultiShot FlexPlate Mach1 T1R Competent Cells are fast growing. Mach1 colonies are clearly visible within eight hours of plating the transformation mix, enabling you to plate and pick colonies the same day. Perform mini-preps after only four hours of growth from an overnight colony—saving an entire day in your experiments. The Mach1 T1R strain is useful for many cloning applications and provides transformation efficiencies of >1 x 108 cfu/µg with control plasmid DNA.

Key features of the MultiShot FlexPlate Mach1 T1R Competent Cells include:
endA1 for cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by endonuclease I
recA1398 for reduced occurrence of non-specific recombination in cloned DNA
hsdR for efficient transformation of unmethylated DNA from PCR amplifications
tonA (fhuA) for T1 and T5 phage resistance
lacZΔM15 for blue/white color screening of recombinant clones

MultiShot StripWell format—flexible for no waste
Each StripWell of eight connected tubes is designed with the same single-tube, single-use features as our One Shot format tubes. This format allows for all steps of the transformation protocol, up to plating, to take place in the same tube, thereby saving time and preventing contamination. After addition of DNA, MultiShot StripWell competent cells can be transformed by heat-shock at 42°C using a water bath, all in the same tube.

The StripWell tubes can be cut to permit transformations in a single tube, eliminating wasted aliquots of cells and avoiding freeze-thaws that can result in reduced transformation efficiency.

Key features of the MultiShot StripWell format include:
Increased productivity—for medium throughput experimentation
Maximal flexibility—transform as few as 1 tube and as many as 96 from the StripWell rack
Familiar convenience—no aliquotting, add DNA directly to cells, heat shock, and recover in the same tube
Peace of mind—single-use transformation minimizes contamination

Genotype: F- φ80(lacZ)ΔM15 ΔlacX74 hsdR(rK-mK+) ΔrecA1398 endA1 tonA

Find the strain and format that you need
We offer many other E. coli strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs. If you require other high-throughput transformation options, choose from our collection of MultiShot formatted comp cells or email us for custom formatting options.

MAX Efficiency™ DH10B Competent Cells (Invitrogen™)

MAX Efficiency DH10B Competent Cells are highly efficient E. coli, perfect for most applications. DH10B E. coli are available in both electrocompetent and chemically competent formats. The high transformation efficiency makes them ideal for generating cDNA or genomic libraries. High-efficiency transformation is seen with plasmids 150 kb in size. The versatile DH10B genotype has the following features:

lacZ for blue/white screening of recombinant clones
• Elimination of mcrA, mcrBC, mrr, and hsdRMS restriction systems to allow construction of more representative genomic libraries
• A1 mutation to increase plasmid yield and quantity

One Shot™ INV110 Chemically Competent E. coli (Invitrogen™)

INV110 E. coli are derived from JM110 E. coli and are specifically designed for the growth and purification of plasmid DNA that will be digested with dam or dcm methylation-sensitive restriction enzymes. INV110 is a significant improvement over the JM110 strain. INV110 has been engineered to carry an endA1 mutation, which eliminates the non-specific endonuclease Endonuclease I, for improved plasmid DNA preparation. In addition, the INV110 genotype features:

• Blue/white screening of recombinant clones (lacZΔM15)
• Reduced cleavage of foreign methylated DNA for improved transformation of genomic DNA [Δ(mcrC-mrr)]
• F´ episome for phage infection and rescue of single-strand DNA

One Shot™ TOP10F´ Chemically Competent E. coli (Invitrogen™)

One Shot® TOP10F´ Chemically Competent E. coli are identical to TOP10 cells, with the addition of an F´ episome. TOP10 competent cells are provided at a transformation efficiency of 1 x 109 cfu/µg supercoiled DNA and are ideal for high-efficiency cloning and plasmid propagation.

Using One Shot® TOP10F´ Chemically Competent E. coli
The F´ episome carries the tetracycline resistance gene and allows isolation of single-stranded DNA from vectors that have an f1 origin of replication. In addition, the F´ episome carries the lacIq repressor for inducible expression from trc, tac, and lac promoters using IPTG. TOP10F´ cells require IPTG induction for blue/white color screening.

One Shot™ ccdB Survival™ 2 T1R Competent Cells (Invitrogen™)

One Shot® ccdB Survival™ 2 T1R Chemically Competent Cells are suitable for the propagation of plasmids containing the ccdB gene, and are designed for use with the Gateway® Vector Conversion System and for propagating Gateway® destination, donor, and supercoiled entry vectors. Transformation efficiency is >1 × 109 transformants/µg plasmid DNA. The ccdB Survival™ 2 T1R E. coli strain is derived from the TOP10 strain. The ccdB Survival™ 2 T1R E. coli strain offers the following features:

• Resistance to the ccdB gene product
• Resistance to T1 and T5 phage (the tonA/fhuA phenotype)
• Supports high-yield preparation of supercoiled plasmid DNA (the endA1 phenotype)
• Reduced nonspecific recombination of cloned DNA (the recA1 phenotype)

Genotype: F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG fhuA::IS2

TOP10F´ Electrocomp™ Kit (Invitrogen™)

The TOP10F' genotype is identical to TOP10 with the addition of an F' episome. The F' episome carries the tetracycline resistance gene and allows isolation of single-stranded DNA from vectors that have an f1 origin of replication. In addition, the F' carries the lacIq repressor for inducible expression from trc, tac, and lac promoters using IPTG. TOP10F' require IPTG induction for blue/white color screening.

The TOP10F' genotype offers the following features:

hsdR for efficient transformation of unmethylated DNA from PCR amplifications
mcrA for efficient transformation of methylated DNA from genomic preparations
lacZΔM15 for blue/white color screening of recombinant clones
endA1 for cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by Endonuclease I
recA1 for reduced occurrence of non-specific recombination in cloned DNA
lacIq repressor for inducible expression from trc, tac, and lac promoters

Note: This kit contains enough electrocompetent cells for 25 x 40 µL rxns or 50 x 20 µL rxns. Please see the product manual for reaction scale recommendations.

Genotype:
F'{lacIqTn10(TetR)} mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL endA1 nupG

MegaX DH10B T1R Electrocomp™ Cells (Invitrogen™)

MegaX DH10B T1R Electrocomp Cells are the highest-efficiency electrocompetent cells available. Improved production processes now guarantee a three-fold greater number of colonies per transformation (>3 x 1010 cfu/µg of pUC DNA), making the MegaX DH10B T1R cells ideal for highly demanding cloning and library construction. They have the same genotype as the widely used DH10B T1R strain, including tonA, which prevents infection by T1 and T5 lytic bacteriophages, and safeguards your valuable clones and libraries.

Using MegaX DH10B T1R cells
MegaX DH10B T1R cells provide highly efficient transformation of methylated DNA, since they lack the E. coli K12 restriction systems (mcrA Δ(mrr-hsdRMS-mcrBC)). The Φ80lacZΔM15 marker provides α-complementation of the β-galactosidase gene, allowing blue/white screening on agar plates containing X-gal or Bluo-gal.

Genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC), Φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL nupG tonA

MAX Efficiency™ DH5α-T1R Competent Cells (Invitrogen™)

MAX Efficiency DH5α T1 Phage-Resistant Competent Cells are high efficiency chemically competent cells for generating cDNA libraries. In addition to supporting blue/white screening, recA1 and endA1 mutations in DH5α increase insert stability and improve the quality of plasmid DNA prepared from minipreps. MAX Efficiency DH5α T1 Phage-Resistant Competent Cells yield >1 × 109 transformants/µg of monomer pUC19 DNA in a 100-µl transformation reaction.

One Shot™ MAX Efficiency™ DH5α-T1R Competent Cells (Invitrogen™)

One Shot MAX Efficiency DH5α-T1R Competent Cells are derived from the popular DH5α strain. DH5α-T1R carry the tonA genotype that confers resistance to T1 and T5 phage. T1 bacteriophage spread rapidly and lyse E. coli hosts, which are commonly used for cloning and library construction. They offer added security to protect valuable clones and libraries. This is especially important for genome and sequencing centers where T1 phage infection would be catastrophic. In addition to phage resistance, DH5α-T1R E. coli offer the following benefits:

• Cleaner preparations of DNA and better results in downstream applications due to the elimination of nonspecific digestion by Endonuclease I (endA1)
• Reduced occurrence of unwanted recombination in cloned DNA (recA1)
• Efficient transformation of unmethylated DNA from PCR applications (hsdR)
• Blue/white color screening of recombinant clones carrying the alpha fragment of β-galactosidase (lacZΔM15)

Convenient and efficient format
DH5α-T1R E. coli are supplied in the convenient, single-reaction One Shot format. Each tube contains enough cells for one transformation so there are no efficiency-zapping, freeze-thaw cycles or money wasted on unused cells.

ElectroMAX™ Stbl4™ Competent Cells (Invitrogen™)

ElectroMAX™ Stbl4™ Competent Cells are specifically designed for cloning unstable inserts. As electrocompetent cells, they have one of the highest transformation efficiencies available (>5×109 cfu/µg), making them ideal for generating cDNA and genomic libraries and for cloning unstable inserts. These electrocompetent E. coli:

• Are ideal for cloning unstable DNA—stabilizes direct repeat and retroviral sequences
• Permit efficient cloning of methylated genomic DNA
• Support blue/white screening
• Are designed to deliver high-yield plasmid preparations for downstream applications
• Provide transformation efficiencies of >5×109 cfu/µg

Propagating unstable and large DNA with high transformation-efficiency cells
Many competent cell strains have the recA1 genotype, which reduces recombination. However there are some instances when the DNA that you are trying to clone is still unstable in such cells, perhaps due to the presence of inverted or direct repeats, or GC-rich tracts. While such sequences are relatively common in eukaryotic genomes, they are rare in E. coli. Consequently, rearrangements may occur when these sequences are introduced into standard E. coli strains.

Using ElectroMAX™ Stbl4™ Cells
ElectroMAX™ Stbl4™ electrocompetent E. coli cells are a derivative of Stbl2™ cells and are ideal for the cloning of unstable inserts such as retroviral sequences, direct repeats, and tandem array genes. Furthermore, ElectroMAX™ Stbl4™ cells are useful for the generation of cDNA libraries using plasmid-derived vectors and are able to take up and maintain large plasmids (e.g., 50 kb cosmids and 100–200 kb P1 clones). The Stbl4™ cells also contain an F' episome, allowing them to serve as a host for single-stranded DNA such as M13mp cloning vectors. The lacZΔM15 marker provides α-complementation of the β-galactosidase gene from pUC or similar vectors, and can therefore be used for blue/white screening of colonies on agar plates containing Xgal or Bluo-gal and IPTG. The mcrA mutation and the mcrBC-hsdRMS-mrr deletion allow cloning of genomic sequences which are methylated. Finally, the endA1 mutation greatly increases plasmid yield and quality.

Genotype: mcrA Δ(mcrBC-hsdRMS-mrr) recA1 endA1 gyrA96 gal-thi-1 supE44 λ-relA1 Δ(lac-proAB)/F' proAB+lacIqZΔM15 Tn10 (TetR)

One Shot™ Stbl3™ Chemically Competent E. coli (Invitrogen™)

One Shot® Stbl3™ Chemically Competent E. coli are designed especially for cloning direct repeats found in lentiviral expression vectors. Unlike TOP10 E. coli, these cells reduce the frequency of homologous recombination of long terminal repeats (LTRs) found in ViraPower™ Lentiviral Expression Vectors and other retroviral vectors. The DNA yield from Stbl3™ cells is often >10-fold higher than alternative strains such as TOP10 (see table). One Shot® Stbl3™ Chemically Competent Cells deliver:

• Optimal performance for the propagation of lentiviral vectors
• Enhanced genomic cloning capacity (mcrB, mrr)
• Expected efficiency of >1 × 108 transformants/µg plasmid DNA
• High DNA yields

Recommended use
The Stbl3™ E. coli strain is derived from the HB101 E. coli strain and is recommended for use when cloning unstable inserts such as lentiviral DNA containing direct repeats (e.g., Invitrogen® ViraPower™ Lentiviral Expression kits). The transformation efficiency of One Shot® Stbl3™ chemically competent cells is greater than 1 × 108 cfu/µg plasmid DNA.

Genotype: F-mcrB mrrhsdS20(rB-, mB-) recA13 supE44 ara-14 galK2 lacY1 proA2 rpsL20(StrR) xyl-5 λ-leumtl-1

Find the strain and format that you need
We offer a variety of competent cells for the propagation of unstable DNA inserts.

MAX Efficiency® Stbl2™ Competent Cells—stable cloning of direct repeat and retroviral sequences and tandem array genes (5 × 200 µL) (>1 × 109 cfu/µg plasmid DNA)
One Shot® Stbl3™ Chemically Competent E. coli—cloning unstable inserts containing direct repeats (20 × 50 µL) (>1 × 108 cfu/µg plasmid DNA)
ElectroMAX™ Stbl4™ Competent Cells—high transformation efficiencies (>5 × 109 cfu/µg plasmid DNA), ideal for generating cDNA and genomic libraries and for cloning unstable inserts