One Shot™ Mach1™ T1 Phage-Resistant Chemically Competent E. coli
One Shot&trade; Mach1&trade; T1 Phage-Resistant Chemically Competent <i>E. coli</i>
Invitrogen™

One Shot™ Mach1™ T1 Phage-Resistant Chemically Competent E. coli

One Shot Mach1 T1 Phage-Resistant Chemically Competent E. coli are the fastest growing chemically competent cells with a doubling timeRead more
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Catalog number C862003
Price (USD)
538.00
Each
Add to cart
Price (USD)
538.00
Each
Add to cart

One Shot Mach1 T1 Phage-Resistant Chemically Competent E. coli are the fastest growing chemically competent cells with a doubling time approximately 50 minutes. Mach1 T1R colonies are clearly visible within eight hours of plating the transformation mix (ampicillin selection), enabling you to plate and pick colonies the same day. Plasmid DNA can be prepared four hours after inoculating a single, overnight-grown colony in the selective media of choice (with any antibiotic). Mach1 T1R Chemically Competent E. coli cells can achieve transformation efficiency of >1 x 109 cfu/μg plasmid DNA.

The Mach1 T1R strain is derived from a non-K-12, wild-type W E. coli strain (ATCC #9637) that had been selected due it’s growth speed and genetically manipulated to possess the most useful cloning strain features. The strain was engineered to carry the tonA (also known as fhuA) mutation that confers resistance to T1 and T5 phage infection. The tonA genotype offers added security to valuable clones and libraries. The Mach1 T1R strain has the lacZΔM15 genotype, allowing blue-white screening on plates containing either X-Gal or Bluo-Gal. It also carries the recA1398 mutation that helps reduce the rate of recombination while propagating plasmid DNA, the endA1 mutation that increases plasmid DNA yield and quantity, and hsdR restriction system deletion to allow genomic DNA cloning.

One Shot Mach1 T1 Phage-Resistant Chemically Competent E. coli offer:
• Transformation efficiencies of >1 x 109 cfu/μg plasmid DNA
• tonA (fhuA) genotype to confer resistance to T1 and T5 phage 
• lacZ∆M15 for blue-white color screening of recombinants
• hsdR mutation for efficient transformation of unmethylated DNA from PCR applications
• recA1398 mutation for reduced occurrence of homologous recombination in cloned DNA
• endA1 mutation for increased plasmid yield and quality

Easy-to-use One Shot format 
Mach1 T1 Phage-Resistant Chemically Competent E. coli cells are supplied in the convenient, single-reaction One Shot format. The single-tube, single-use format allows all steps of the transformation protocol, up to plating, to take place in the same tube, helping save time and prevent contamination.

Genotype
F– φ80lacZΔM15 ΔlacX74 hsdR(rK–, mK+) ΔrecA1398 endA1 tonA

Find the strain and format that you need 
We offer other strains in chemically competent and electrocompetent cell formats to meet your specific needs.
Mach1 T1R and other strains are available in MultiShot formats for high throughput applications.
Explore bacterial growth media formats.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeChemically Competent Cells
Contains F' EpisomeNo
Improves Plasmid QualityYes (endA1)
Cloning Methylated DNANo
Transformation Efficiency LevelHigh Efficiency (>1 x 109 cfu/μg)
Antibiotic Resistance BacterialNo
Cloning Unstable DNANot suitable for cloning unstable DNA
Blue/White ScreeningYes (lacZΔM15)
High-throughput CompatibilityLow
Preparing Unmethylated DNANo
Reduces RecombinationYes (recA1)
Shipping ConditionDry Ice
T1 Phage - Resistant (tonA)Yes
SpeciesE. coli (wild-type W)
FormatTube
Product LineOne Shot™
Quantity21 x 50 μL/tube
Unit SizeEach
Contents & Storage
• One Shot Mach1 T1R Chemically Competent E. coli (21 x 50 μL); store at –80°C
• pUC19 DNA (50 μL at 10 pg/μL); store at –20°C
• S.O.C. Medium (6 mL); store at 4°C or room temperature

Frequently asked questions (FAQs)

I'm making a yeast genomic library and want to transform and amplify it in one of your competent cell strains. What genotype features should I look for in choosing a good strain?

We would recommend a mcr/mrr- strain, which prevents restriction of methylated eukaryotic DNA in the E. coli host. We would also recommend using a T1R strain, as T1 is a common contaminant in genomic/cDNA libraries.

Which is your fastest growing strain of competent cells?

Our Mach1-T1R competent cells grow faster than any of our common cloning strains. It has a doubling time of 54 minutes versus doubling times in excess of 70 mins for standard cloning strains, such as DH5α cells. Colonies of Mach1-T1R begin to be visible on a plate 8 hours after plating the transformation mix at 37 degrees C. It can be mini-prepped from 1.5 mL cultures in as little as 4 hours at 37 degrees C after inoculation with a single large overnight colony.

What advantages do your Stbl2 cells offer over other cloning strains?

There are other strains available that may function similarly to Stbl2 cells in stabilizing inserts or vectors with repeated DNA sequences. However, one advantage of Stbl2 cells over many similar strains is that they are sensitive to Kanamycin, so you can use Stbl2 to propagate plasmids containing a Kanamycin resistance marker. 

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.

Is S.O.C. medium absolutely required when recovering competent bacterial cells during transformation?

Many media can be used to grow transformed cells, including standard LB, SOB or TB broths. However, S.O.C. is the optimal choice for recovery of the cells before plating. The nutrient-rich formula with added glucose is often important for obtaining maximum transformation efficiencies.

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