ElectroMAX™ DH10B T1 Phage-Resistant Competent Cells
ElectroMAX™ DH10B T1 Phage-Resistant Competent Cells
Invitrogen™

ElectroMAX™ DH10B T1 Phage-Resistant Competent Cells

ElectroMAX DH10B T1 Phage-Resistant Competent Cells offer transformation efficiencies that are among the highest we offer and can exceed 1Read more
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Catalog number 12033015
Price (USD)
387.00
Each
Add to cart
Price (USD)
387.00
Each
Add to cart

ElectroMAX DH10B T1 Phage-Resistant Competent Cells offer transformation efficiencies that are among the highest we offer and can exceed 1 x 1010 cfu/μg plasmid DNA in efficiency. They are ideal for applications requiring high-efficiency transformation, including cDNA and gDNA library construction.

ElectroMAX DH10B cells:
• Maximize yield of transformants from limited cloning products
• Permit efficient cloning of methylated DNA
• Are resistant to T1 and T5 bacteriophages
• Support blue/white screening by α-complementation on plates containing X-Gal or Bluo-Gal
• Deliver high-yield plasmid preparations for downstream applications

Phage-resistant representative libraries from a single transformation
The tonA mutation in DH10B cells confers resistance to bacteriophages T1 and T5, safeguarding valuable libraries against contamination. Additionally, mutations in the methylation-dependent restriction system (mcrA, mcrBC, and mrr) make ElectroMAX DH10B T1 Phage-Resistant Cells ideal for construction of genomic libraries of both prokaryotic and eukaryotic genomic DNA and allow for efficient plasmid rescue from eukaryotic genomes. ElectroMAX DH10B T1 Phage-Resistant Cells are suitable for construction of gene banks, for the generation of cDNA libraries using plasmid-derived vectors, and for situations when DNA is limiting. This strain also has the Φ80lacZΔM15 genotype, providing for the option of blue/white screening on plates containing either X-Gal or Bluo-Gal. Finally, the endA1 mutation makes DH10B an excellent vehicle for amplifying plasmid DNA for subsequent extraction and purification.

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

Find the strain and format that fits your needs
DH10B cells are available in both electrocompetent and chemically competent formats.
ElectroMAX DH10B cells are suitable for cDNA or gDNA library construction but are not resistant to T1 and T5 phage infection.
MegaX DH10B T1R Electrocomp Cells have the highest transformation efficiency (>3 x 1010 cfu/μg plasmid DNA) and also offer the benefit of T1 and T5 phage resistance.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeElectrocompetent Cells
Contains F' EpisomeNo
Improves Plasmid QualityYes (endA1)
Cloning Methylated DNAYes (mcrA)
Transformation Efficiency LevelHigh Efficiency (>1 x 109 cfu/μg)
Antibiotic Resistance BacterialYes (Streptomycin)
Cloning Unstable DNANot suitable for cloning unstable DNA
Blue/White ScreeningYes (lacZΔM15)
High-throughput CompatibilityLow
PlasmidMay be used for plasmids >20 kb
Preparing Unmethylated DNANo
Reduces RecombinationYes (recA1)
Shipping ConditionDry Ice
T1 Phage - Resistant (tonA)Yes
SpeciesE. coli (K12)
FormatTube
Product LineElectroMAX™
Quantity5 x 100 μL
Unit SizeEach
Contents & Storage
• ElectroMAX DH10B T1 Phage-Resistant Cells (5 x 100 μL)
Store Competent Cells at –80°C.

• pUC19 DNA (50 μL at 10 pg/μL)
Store pUC19 DNA at –20°C.

• S.O.C. Medium (2 x 6 mL)
Store S.O.C. Medium at 4°C or room temperature.

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Frequently asked questions (FAQs)

How do you recommend that I prepare my DNA for successful electroporation of E. coli?

For best results, DNA used in electroporation must have a very low ionic strength and a high resistance. A high-salt DNA sample may be purified by either ethanol precipitation or dialysis.

The following suggested protocols are for ligation reactions of 20ul. The volumes may be adjusted to suit the amount being prepared.

Purifying DNA by Precipitation: Add 5 to 10 ug of tRNA to a 20ul ligation reaction. Adjust the solution to 2.5 M in ammonium acetate using a 7.5 M ammonium acetate stock solution. Mix well. Add two volumes of 100 % ethanol. Centrifuge at 12,000 x g for 15 min at 4C. Remove the supernatant with a micropipet. Wash the pellet with 60ul of 70% ethanol. Centrifuge at 12,000 x g for 15 min at room temperature. Remove the supernatant with a micropipet. Air dry the pellet. Resuspend the DNA in 0.5X TE buffer [5 mM Tris-HCl, 0.5 mM EDTA (pH 7.5)] to a concentration of 10 ng/ul of DNA. Use 1 ul per transformation of 20 ul of cell suspension.

Purifying DNA by Microdialysis: Float a Millipore filter, type VS 0.025 um, on a pool of 0.5X TE buffer (or 10% glycerol) in a small plastic container. Place 20ul of the DNA solution as a drop on top of the filter. Incubate at room temperature for several hours. Withdraw the DNA drop from the filter and place it in a polypropylene microcentrifuge tube. Use 1ul of this DNA for each electrotransformation reaction.

When should DMSO, formamide, glycerol and other cosolvents be used in PCR?

Cosolvents may be used when there is a failure of amplification, either because the template contains stable hairpin-loops or the region of amplification is GC-rich. Keep in mind that all of these cosolvents have the effect of lowering enzyme activity, which will decrease amplification yield. For more information see P Landre et al (1995). The use of co-solvents to enhance amplification by the polymerase chain reaction. In: PCR Strategies, edited by MA Innis, DH Gelfand, JJ Sninsky. Academic Press, San Diego, CA, pp. 3-16.

Additionally, when amplifying very long PCR fragments (greater than 5 kb) the use of cosolvents is often recommended to help compensate for the increased melting temperature of these fragments.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.