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D4154

Sigma-Aldrich

Plasmid DNA from E. coli RRI

pUC18, buffered aqueous solution

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About This Item

CAS Number:
9007-49-2
MDL number:
MFCD00166896
UNSPSC Code:
41105600
NACRES:
NA.85
Pricing and availability is not currently available.

biological source

Escherichia coli (RRI)

Quality Level

200

grade

for molecular biology

form

buffered aqueous solution

shipped in

dry ice

storage temp.

−20°C

SMILES string

[P](=O)(OC2C(OC(C2)N)CO[P](=O)(OC3C(OC(C3)N)CO)O)(OCC1OC(CC1O)N)O

InChI

1S/C15H31N3O13P2/c16-13-1-7(20)11(28-13)5-25-32(21,22)31-9-3-15(18)29-12(9)6-26-33(23,24)30-8-2-14(17)27-10(8)4-19/h7-15,19-20H,1-6,16-18H2,(H,21,22)(H,23,24)

InChI key

AWBASQCACWFTGD-UHFFFAOYSA-N

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General description

Plasmid DNA from Escherichia coli RRI confers 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.

Application

Plasmid DNA from Escherichia coli RRI has been used in polymerase chain reaction (PCR).[1]
Plasmid DNA from E. coli RRI has also been used for in vitro DNA transfection studies.[2]

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.

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.

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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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Certificates of Analysis (COA)

Lot/Batch Number
SLCB5238
SLBZ8650
016K61551
055K61072
055K61071

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Questions

1–2 of 2 Questions  

  1. What is the sequence of plasmid pUC18 (product number D4154) and the number of copies of the plasmid contained in this product? It will be used as an internal control for PCR amplification.

    What is the sequence of plasmid pUC18 (product number D4154) and the number of copies of the plasmid contained in this product? It will be used as an internal control for PCR amplification.

    1 answer

    1. The GenBank accession number for the sequence of the high-copy number plasmid pUC18 is VB0025.

      The following link contains the sequence for this plasmid: https://www.addgene.org/50004/sequences/#depositor-full.

      The plasmid is 2,686 base pairs in length and the molecular weight is approximately 1.75 × 10⁶ daltons, so the 10µg should be 3.4 x 10^12 copies.

      Helpful?

  2. user manual for Puc18?

    1 answer

    1. The Puc18 is not supplied with a user manual or specific protocol. Please see the link below to the product data sheet which provides additional information on this plasmid: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/263/858/d4154dat.pdf

      Helpful?

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