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D4154 Sigma

Plasmid DNA from E. coli RRI

pUC18, buffered aqueous solution

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Properties

grade   for molecular biology
form   buffered aqueous solution
shipped in   dry ice
storage temp.   −20°C

Description

Components

pUC18 Plasmid DNA, 10 μg is supplied at approximately 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0) with 1 mM EDTA.

Biochem/physiol Actions

These plasmids confer ampicillin resistance and complement defects in β-galactosidase in appropriate host strains. The multiple cloning site (MCS) is within the β-galactosidase gene; pUC8 and 9 have nine unique sites within the MCS while pUC18 and 19 have thirteen.
Foreign DNA inserted at the MCS abolishes the ability to catabolize lactose. Lactose-positive, ampicillin-resistant colonies (host strain containing plasmid) form blue colonies on plates containing ampicillin and X-Gal; lactose-negative, ampicillin-resistant colonies (host strain containing plasmid with foreign DNA inserted at the MCS) form white colonies on this medium. The orientations of the MCS regions in the pUC plasmids are analogous to those of the corresponding M13 phage.

Principle

Foreign DNA inserted at the MCS interrupts the β-galactosidase gene and abolishes the ability to catabolize lactose. Lactose-positive, ampicillin-resistant colonies (host strain containing plasmid) form blue colonies on plates containing ampicillin and X-Gal; lactose-negative, ampicillin-resistant colonies (host strain containing plasmid with foreign DNA inserted at the MCS) form white colonies on this medium.

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Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
Protocols & Articles

Articles

Introduction to Blue-White Screening

Blue-white screening is a rapid and efficient technique for the identification of recombinant bacteria. It relies on the activity of β-galactosidase, an enzyme occurring in E. coli, which cleaves lac...
Keywords: Antibiotics, Cloning, Culture media, Gene expression, Immunocytochemistry, Peptide synthesis, Polymerase chain reaction, Recombination, Sterilizations, transformation

Peer-Reviewed Papers
15

References

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