Invitrogen™ One Shot™ TOP10 Chemically Competent E. coli Cells

One Shot™ TOP10 Chemically Competent E. coli are designed for high-efficiency cloning and plasmid DNA propagation. These cells are supplied with a transformation efficiency of >1 x 10⁹ cfu/μg plasmid DNA, making them ideal for use in various molecular biology applications, such as genomic DNA cloning, plasmid rescue, and high-copy plasmid propagation. TOP10 cells are genetically similar to the DH10B strain but are more resilient to stress conditions like osmotic shock and acidic pH stress.

These cells are preparation ready for use with blue/white screening systems and are suitable for cloning both prokaryotic and eukaryotic genomic DNA. Their design ensures they deliver enhanced stability and better results in plasmid DNA preparation.

Key Features & Benefits:

1. High Transformation Efficiency:
   • With a transformation efficiency of >1 x 10⁹ cfu/μg, these cells are suitable for applications requiring high-efficiency DNA cloning, such as plasmid cloning, library construction, and plasmid preparation.
2. Genetic Modifications for Stability:
   • recA1 mutation reduces the occurrence of nonspecific recombination in cloned DNA, ensuring the stability of inserted sequences.
   • endA1 mutation improves plasmid DNA quality by eliminating endonuclease I activity, which would otherwise lead to nonspecific digestion of the plasmid.
3. Cloning of Both Methylated and Unmethylated DNA:
   • The mcrA mutation allows for the cloning of methylated DNA, which is important for cloning genomic DNA from eukaryotic sources that may have naturally occurring methylated DNA sequences.
   • hsdR mutation ensures efficient transformation of unmethylated DNA from PCR amplifications.
4. Blue/White Screening:
   • lacZΔM15 genotype allows for blue/white colony screening on agar plates containing X-Gal or Bluo-Gal, simplifying the identification of recombinant clones.
5. Single-Tube One Shot Format:
   • The One Shot format allows for easy-to-use, single-reaction transformations where all steps from DNA addition to plating occur in the same tube. This reduces handling time and minimizes the risk of contamination, especially in high-throughput applications.
6. Ideal for High-Copy Plasmid Propagation:
   • These cells are particularly useful for the propagation of high-copy plasmids, ensuring high yield of plasmid DNA for subsequent applications, such as gene expression or cloning.

Applications:

1. Routine Cloning Experiments:
   • One Shot TOP10 cells are ideal for cloning plasmids, whether you are working with eukaryotic genomic DNA or PCR products.
2. Plasmid DNA Propagation:
   • Used extensively for propagating plasmids in high-copy number for downstream applications, including protein expression and genetic studies.
3. Plasmid Rescue:
   • These cells are useful for rescuing plasmids from eukaryotic genomes in order to clone large fragments of eukaryotic DNA efficiently.
4. Library Construction:
   • Perfect for generating cDNA libraries or gene libraries from both prokaryotic and eukaryotic sources, enabling the study of a wide range of genetic material.
5. Site-Directed Mutagenesis:
   • Can be used in mutagenesis experiments to generate targeted mutations in plasmid sequences, allowing for genetic manipulation.

Genotype:

• F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK λ– rpsL(StrR) endA1 nupG

Specifications:

Contents:

• 10 x 50 μL DH10B Competent Cells
• pUC19 DNA (50 μL, 10 pg/μL)
• S.O.C. Medium (5 mL)

Storage:

• Store the competent cells at -80°C for long-term storage. The cells should be thawed gently on ice before use.

Thermo Scientific™ One Shot™ TOP10 Chemically Competent E. coli are high-efficiency, chemically competent cells perfect for routine cloning, high-copy plasmid propagation, and cDNA library construction. With features like blue/white screening, methylated DNA cloning, and a convenient One Shot format, they offer enhanced cloning performance, stability, and ease of use in molecular biology workflows.