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DMT Competent Cell (DpnI+) – CD511

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DMT Chemically Competent Cell


Characteristics of DMT Competent Cell

  • High transformation efficiency: >10ˆ8 cfu/μg (pUC19 DNA).
  • Resistance to T1 and T5 phage.
  • Expresses DMT enzyme (DpnI) to achieve in vivo digestion of methylated DNA
  • Ideal for site-directed mutagenesis if high-fidelity PCR was performed on a methylated DNA template or vector.

Genotype of DMT Competent Cell (DpnI+)

F- φ80 lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK-, mK+) phoA supE44 thi -1 gyrA96 relA1 tonA


Product Manual and Datasheet:

CD511 DMT Chemically Competent Cell


Genotype definitions (if applicable to this product)

Genotype Description
ara-14 Blocks arabinose catabolism
argF Ornithine carbamoyltransferase mutation blocks ability to use arginine
dam/dcm Abolishes endogenous adenine methylation at GATC sequences (dam) or cytosine methylation at CCWGG sequences (dcm). Used to propagate DNA for cleavage with certain restriction enzymes (e.g. Ava II, Bcl I)
DE3 Lysogen that encodes T7 RNA polymerase. Used to induce expression in T7-driven expression systems
endA endA Mutation in the non-specific endonuclease Endonuclease I; eliminates non-specific endonuclease activity, resulting in improved plasmid preps
A self-transmissible, low-copy plasmid used for the generation of single-stranded DNA when infected with M13 phage; may contain a resistance marker to allow maintenance and will often carry the lacI and lacZ∆M15 genotypes
galK Galactokinase mutation blocks catabolism of galactose—cells that are galK minus grow in the presence of galactose as the sole carbon source
galU Glucose-1-phosphate uridylyltransferase mutation blocks ability to use galactose—cells that are galU minus can grow on media that contains galactose as the sole carbon source
gyrA96 DNA gyrase mutant produces resistance to nalidixic acid
hsd Mutations in the system of methylation and restriction that allow E. coli to recognize DNA as foreign. The hsd genotype allows efficient transformation of DNA generated from PCR reactions *hsdR–eliminates restriction of unmethylated EcoK I sites. (1) **hs
lacI Encodes the lac repressor that controls expression from promoters that carry the lac operator; IPTG binds the lac repressor and derepresses the promoter; often used when performing blue/white screening or to control expression of recombinant genes
lacY1 Blocks use of lactose via β-D-galactosidase mutant
lacZ β-D-galactosidase gene; mutations yield colorless (vs. blue) colonies in the presence of X-gal
lacZ∆M15 Element required for β-galactosidase complementation when plated on X-gal; used in blue/white screening of recombinants; usually carried on the lambdoid prophage φ80 or F´
leuB Requires leucine for growth on minimal media via β-isopropyl malate dehydrogenase mutation
lon lon Deficiency in the Lon ATPase-dependent protease; decreases the degradation of recombinant proteins; all B strains carry this mutation
mcrA, mcrBC,or mrr Mutations that allow methylated DNA to not be recognized as foreign; this genotype is necessary when cloning genomic DNA or methylated cDNA
nupG Mutation for the transport of nucleosides
ompT Indicates that the E. coli lack an outer membrane protease—reduces degradation of heterologous the strains and recovery of intact recombinant proteins is improved in ompT minus strains
P3 A 60-kb low-copy plasmid that carries the ampicillin and tetracycline resistance genes with amber mutations; used predominantly for selection of supF-containing plasmids; carries the kanamycin resistance gene for selection
pLys pLys Plasmid that encodes T7 lysozyme; used to reduce basal expression in T7-driven expression systems by inhibiting basal levels of T7 RNA polymerase
proAB proAB Requires proline for growth on minimal media
recA Mutation in a gene responsible for general recombination of DNA; particularly desirable when cloning genes with direct repeats
relA RNA is synthesized in absence of protein synthesis (relaxed phenotype) relA locus regulates the coupling between transcription and translation. In the wild type, limiting amino acid concentrations results in the shutdown of RNA synthesis (also known as th
rpsL Confers resistance to streptomycin (this makes a mutant ribosomal protein, small subunit, the target of the drug)
supE,F tRNA glutamine suppressor of amber (supE)(UAG) or tyrosine (supF)
thi-1 Requires thiamine for growth on minimal media
Tn10 Confers tetracycline resistance via a transposon
tonA Confers resistance to the lytic bacteriophage T1, T5 and f80
traD, D36 Prevents transfer of F’ episome via transfer factor mutation
tsx Confers resistance to phage T6 and colicin K
xyl-5 Blocks catabolism of xylose

Additional information

Format

10 x 50 ul, 20 x 50 ul

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