Store Competent Cells at -80°C. Store pUC19 DNA at
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ElectroMAX™ Stbl4™ Competent Cells are specifically designed for cloning unstable inserts. As electrocompetent cells, they have one of the highest transformation efficiencies available (>5×109cfu/µg), making them ideal for generating cDNA and genomic libraries and for cloning unstable inserts. These electrocompetentE. coli:
• Are ideal for cloning unstable DNA—stabilizes direct repeat and retroviral sequences • Permit efficient cloning of methylated genomic DNA • Support blue/white screening • Are designed to deliver high-yield plasmid preparations for downstream applications • Provide transformation efficiencies of >5×109cfu/µg
Propagating unstable and large DNA with high transformation-efficiency cells Many competent cell strains have therecA1 genotype, which reduces recombination. However there are some instances when the DNA that you are trying to clone is still unstable in such cells, perhaps due to the presence of inverted or direct repeats, or GC-rich tracts. While such sequences are relatively common in eukaryotic genomes, they are rare inE. coli. Consequently, rearrangements may occur when these sequences are introduced into standardE. colistrains.
Using ElectroMAX™ Stbl4™ Cells ElectroMAX™ Stbl4™ electrocompetentE. colicells are a derivative of Stbl2™ cells and are ideal for the cloning of unstable inserts such as retroviral sequences, direct repeats, and tandem array genes. Furthermore, ElectroMAX™ Stbl4™ cells are useful for the generation of cDNA libraries using plasmid-derived vectors and are able to take up and maintain large plasmids (e.g., 50 kb cosmids and 100–200 kb P1 clones). The Stbl4™ cells also contain an F' episome, allowing them to serve as a host for single-stranded DNA such as M13mp cloning vectors. ThelacZΔM15 marker provides α-complementation of the β-galactosidase gene from pUC or similar vectors, and can therefore be used for blue/white screening of colonies on agar plates containing Xgal or Bluo-gal and IPTG. ThemcrA mutation and themcrBC-hsdRMS-mrrdeletion allow cloning of genomic sequences which are methylated. Finally, theendA1 mutation greatly increases plasmid yield and quality.
ElectroMAX™ Stbl4™ Competent Cells are specifically designed for cloning unstable inserts. As electrocompetent cells, they have one of the highest transformation efficiencies available (>5×109cfu/µg), making them ideal for generating cDNA and genomic libraries and for cloning unstable inserts. These electrocompetentE. coli:
• Are ideal for cloning unstable DNA—stabilizes direct repeat and retroviral sequences • Permit efficient cloning of methylated genomic DNA • Support blue/white screening • Are designed to deliver high-yield plasmid preparations for downstream applications • Provide transformation efficiencies of >5×109cfu/µg
Propagating unstable and large DNA with high transformation-efficiency cells Many competent cell strains have therecA1 genotype, which reduces recombination. However there are some instances when the DNA that you are trying to clone is still unstable in such cells, perhaps due to the presence of inverted or direct repeats, or GC-rich tracts. While such sequences are relatively common in eukaryotic genomes, they are rare inE. coli. Consequently, rearrangements may occur when these sequences are introduced into standardE. colistrains.
Using ElectroMAX™ Stbl4™ Cells ElectroMAX™ Stbl4™ electrocompetentE. colicells are a derivative of Stbl2™ cells and are ideal for the cloning of unstable inserts such as retroviral sequences, direct repeats, and tandem array genes. Furthermore, ElectroMAX™ Stbl4™ cells are useful for the generation of cDNA libraries using plasmid-derived vectors and are able to take up and maintain large plasmids (e.g., 50 kb cosmids and 100–200 kb P1 clones). The Stbl4™ cells also contain an F' episome, allowing them to serve as a host for single-stranded DNA such as M13mp cloning vectors. ThelacZΔM15 marker provides α-complementation of the β-galactosidase gene from pUC or similar vectors, and can therefore be used for blue/white screening of colonies on agar plates containing Xgal or Bluo-gal and IPTG. ThemcrA mutation and themcrBC-hsdRMS-mrrdeletion allow cloning of genomic sequences which are methylated. Finally, theendA1 mutation greatly increases plasmid yield and quality.