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CMC0010 Sigma-Aldrich

CHANGER Electrocompetent Cells

for making Uracil-containing DNA for mutagenesis

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Properties

grade   for molecular biology
form   buffered aqueous solution
cell transformation   competent cell type: electrocompetent
transformation efficiency: ~1 × 109 cfu/μg
shipped in   dry ice
storage temp.   −70°C

Description

General description

The CHANGER Electrocompetent Cells are high-efficiency competent cells.
This cell line is ideally suited to the Kunkel directed mutagenesis method.
This method relies on the cells producing plasmid DNA containing some uracil residues. The dut- and ung- mutations prevent the cells from removing uracil from DNA caused by misincorporation or dCTP deamination.
Note that these cells contain an F′ which confers resistance to Chloramphenicol at low levels.

Genotype

[F’ Tra+ Pil+ (CamR)] ung-1 relA1 dut-1 thi-1 spoT1 mcrA

Features and Benefits

• High efficiency cells to create uracil-containing DNA for site-directed mutagenesis
• Sole source of highly efficient Electrocompetent cells (1 × 109 cfu/μg)
• ung- and dut- mutations to generate DNA with some uracil residues
• Ideal genotype and efficiency for Kunkel directed mutagenesis method1.

Components

• CHANGER electrocompetent cells
• pUC 19 transformation control DNA
• recovery medium for cloning

Safety & Documentation

Safety Information

RIDADR 
UN 3245 9

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Competent Cell Selection & E. coli Markers Guide

We offer a range of Escherichia coli bacterial cells made competent with the highest efficiencies by optimized procedure specific to each strain. Choose from 24 new competent cells for a wide variety...
Keywords: Bacterial conjugations, Cloning, Gene expression, Methylations, Recombination, Transcription, transformation

Molecular Biology Handbook

Molecular Biology is the science that aims to understand biological activity at the molecular level. These biological activities usually involve the plant or animal cell, and the nucleic acids and pr...
Keywords: Cell culture, Cell signaling, Cloning, Detection methods, Electrophoresis, Gel electrophoresis, Gene expression, Genetics, Molecular biology, Molecular biology techniques, Nucleic acid hybridization, Nucleic acid purification, Polymerase chain reaction - quantitative, Protein electrophoresis, Purification, Transfection, Western blot, transformation

Protocols

Selecting Correctly Expressing Recombinants

Blue / White colony screening is a strategy to quickly and easily distinguish between recombinant and non-recombinant colonies. It requires a special vector and a special strain of E. coli. It is par...
Keywords: AGE, Amplification, Antibiotics, Cell disruption, Centrifugation, Cloning, Degradations, Detergents, Diagnostic, Digestions, Electrophoresis, Gel electrophoresis, Molecular biology, Peptide synthesis, Phase transitions, Polymerase chain reaction, Precipitation, Purification, Sequencing, Transfection, transformation

Transforming E. coli with Engineered Plasmid

Inoue and colleagues developed this method in 1990. It works well for many strains commonly used in cloning. The original method calls for growing the overnight E. coli cultures at 18°C. We find that...
Keywords: Antibiotics, Centrifugation, Cloning, Peptide synthesis, Phase transitions, transformation

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L3022 LB Broth (Lennox), Highly-referenced microbial growth powder medium, low salt, suitable for salt-sensitive E.coli culture.
L2897 LB Broth with agar (Lennox), Highly-referenced microbial growth powder medium with Agar, low salt, suitable for salt-sensitive E. coli culture.
L5542 LB Agar Plates, pre-poured LB agar plates for E. coli
CMR0002 Recovery Medium for Cloning, for use with competent cells

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