Invitrogen™ One Shot™ INV110 Chemically CompetentE. coli

Description

• One Shot INV110 Chemically Competent E
• coli cells are derived from the JM110 strain and are specifically designed for the growth and purification of plasmid DNA that will be digested with dam or dcm methylation-sensitive restriction enzymes
• The dam and dcm deficiency allows for the production of plasmid DNA that is unmethylated at GmATC and CmC(A/T)GG sites
• INV110 Chemically Competent cells can achieve a transformation efficiency of >1 x 10 6 cfu/μg of control DNA
• The INV110 strain is a significant improvement over the JM110 strain
• The INV110 strain has been engineered to carry an end A1 mutation, which eliminates the non-specific Endonuclease I, for improved plasmid DNA preparation
• Mutation in the methylation-dependent restriction system Δ( mcr C- mrr ) allows transformation of genomic DNA
• This strain also has the lac ZΔM15 genotype, providing for the option of blue-white screening on plates containing either X-Gal or Bluo-Gal
• The INV110 strain contains an F´ episome that can support cloning plasmids with an f1-like origin of replication (phagemids) and allows isolation of single-stranded DNA (ssDNA)
• In addition, the F´ episome carries the lac Iq repressor for inducible expression from trc , tac , and lac promoters using IPTG
• One Shot INV110 Chemically Competent E
• coli offer: • Transformation efficiencies of >1 x 10 6 cfu/μg • dam and dcm deficiency to allow restriction digestion with dam and dcm-sensitive restriction enzymes • Stable F´ episome that allows isolation of ssDNA and carries lac Iq repressor for inducible expression from trc , tac , and lac promoters • Δ( mcr C- mrr ) mutation that reduces cleavage of foreign methylated DNA and improves transformation of genomic DNA • lac ZΔM15 for blue-white color screening of recombinant clones • end A1 for cleaner DNA preparations and better results in downstream applications due to elimination of nonspecific digestion by Endonuclease I • Tn10 to permit selection of the host strain using tetracycline Note: dam deletion results in higher mutation rate, higher recombination frequency, and lower viability on plates
• Avoid picking colonies that are noticeably larger than the rest of the population (potential revertants)
• Pick colonies and isolate the DNA within one day of transformation
• Convenient and efficient format One Shot INV110 Chemically Competent E
• coli cells are supplied in the convenient, single-reaction One Shot format
• Each tube contains enough cells for one transformation, reducing both freeze-thaw cycles and costs associated with unused cells
• Genotype F´ [ tra D36 pro AB lac Iq lac ZΔM15] rps L (StrR) thr leu end A thi -1 lac Y gal K gal T ara ton A tsx dam dcm sup E44 Δ( lac - pro AB) Δ( mcr C- mrr )102:: Tn 10(TetR) Find the strain and format that fits your needs • We offer other strains in chemically competent and electrocompetent cell formats to meet your specific needs
• • Strains in MultiShot formats for high throughput applications are available
• • Alternative strains for ssDNA production are available, such as TOP10F´ (Cat
• No
• C303003) or electrocompetent DH12S (Cat
• No
• 18312017)
• • Explore restriction enzyme details to find methylation sensitivity.