Invitrogen™ TOP10F' Electrocomp™ Kit

Description

TOP10F' Electrocomp Kit E. coli cells are identical to TOP10 cells, with the addition of an F' episome.

TOP10F´ Electrocomp Kit E. coli cells are identical to TOP10 cells, with the addition of an F´ episome. The F´ episome carries the tetracycline resistance gene and allows isolation of single-stranded DNA (ssDNA) 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´ competent cells are transformed via electroporation, thus achieving higher transformation efficiencies than chemically competent cells and may be used in more challenging applications, including cloning a low amount of DNA or cDNA and gDNA library construction. These cells allow stable replication of high copy number plasmids and are provided at a transformation efficiency of >1 x 10¹⁰ cfu/ μg supercoiled DNA and are ideal for high-efficiency cloning and plasmid propagation.

As a derivative of TOP10, TOP10F´ cells carry mutations in the methylation-dependent restriction system (mcrA, mcrBC, and mrr) allowing one to clone both prokaryotic and eukaryotic genomic DNA, as well as efficient plasmid rescue from eukaryotic genomes. Similar to other DH strains, the TOP10 strain also has recA1 and endA1 mutations to increase insert stability and improve the quality of plasmid DNA prepared from minipreps.

TOP10F´ Electrocomp Kit E. coli cells offer:
• Transformation efficiencies of >1 x 10¹⁰ cfu/μg
• Stable F´ episome that allows isolation of ssDNA and carries lacIq repressor for inducible expression from trc, tac, and lac promoters 
• 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 on plates containing either X-Gal or Bluo-Gal
• endA1 for cleaner DNA preparations and better results in downstream applications due to elimination of nonspecific digestion by Endonuclease I
• recA1 for reduced occurrence of nonspecific recombination in cloned DNA

Note: TOP10F´ cells require IPTG induction for blue-white color screening. A high-voltage electroporation apparatus is also required.

Genotype
F´ [lacIq, Tn10(TetR)] mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL(StrR) endA1 nupG

Find the strains and formats that fit your needs
• We offer other DH strains in chemically competent and electrocompetent cell.
Electroporation cuvettes with 0.1 cm (Cat. No. P41050), 0.2 cm (Cat. No. P45050), and 0.4 cm (Cat. No. P46050) gaps are available.
• The TOP10 strain is available in several MultiShot formats for high throughput applications.
• We offer other strains for ssDNA production, such as DH12S (Cat. No. 18312017).

Specifications

• Product Type: Electrocompetent Cells
• Improves Plasmid Quality: Yes (endA1)
• Cloning Methylated DNA: Yes (mcrA)
• Transformation Efficiency Level: High Efficiency (>10¹⁰ cfu/μg)
• Content And Storage: • TOP10F' Electrocomp E. coli (10 x 100 μL)
  Store at -80°C.
  
  • pUC19 vector (50 μL at 10 pg/μL)
  Store pUC19 DNA at -20°C.
  
  • S.O.C. Medium (15 mL)
  Store at room temperature.
  
  
• Cloning Unstable DNA: Not suitable for cloning unstable DNA
• Blue/White Screening: Yes (lacZΔM15)
• Plasmid: May be used for plasmids >20 kb
• Preparing Unmethylated DNA: No
• Reduces Recombination: Yes (recA1)
• Shipping Condition: Dry Ice
• Species: E. coli (K12)
• Format: Tube
• Product Line: Electrocomp™
• Quantity: 10 x 100 μL

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.