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B3928

Sigma-Aldrich

Blue-White Select Screening Reagent

for selection of recombinant bacterial clones

Sinonimo/i:

Blue-White Reagent, Screening Reagent, Selection Reagent

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About This Item

Codice UNSPSC:
12352200
NACRES:
NA.85

Grado

for molecular biology

Livello qualitativo

Stato

liquid

impiego

sufficient for 125 dishes (90 mm)

tecniche

nucleic acid detection: suitable

Compatibilità

suitable for nucleic acid staining

Condizioni di spedizione

dry ice

Temperatura di conservazione

−20°C

Descrizione generale

Sigma′s Blue-White Select Screening Reagent is a chromogenic substrate for β-galactosidase, used to determine the presence or absence of a cloned DNA insert in bacteria growing on agar plates. Blue-White Select Screening Reagent is designed for blue-white selection of recombinant bacterial colonies with the lac+ phenotype.

Caratteristiche e vantaggi

  • Intense color contrast for easy colony selection
  • Convenient, ready-to-use solution

Componenti

40 mg/mL IPTG
40 mg/mL X-Gal
(prepared in DMSO)

Principio

When Blue-White Select Screening Reagent is cleaved by β-galactosidase, the resulting product will form a blue precipitate. Lac+ colonies grown in the presence of Blue-White Select Screening Reagent remain white in color, allowing for easy differentiation between lac+ and lac- colonies.

The technique is based on vectors such as the pUC and the M13mp series that carry a fragment of the β-galactosidase gene encoding an a-fragment of β-galactosidase. Exploitation of these vectors requires use of a bacteria strain carrying the complementing gene fragment to allow the assembly of an active complex, resulting in the formation of blue colonies. Disruption of the ß-galactosidase gene by insertion of a DNA fragment into the vector′s multiple cloning site results in the loss of functional β-galactosidase activity; these colonies remain white, allowing for easy differentiation between lac+ and lac- colonies.

Note legali

Blue-White Select is a trademark of Sigma-Aldrich Co. LLC

Codice della classe di stoccaggio

10 - Combustible liquids

Classe di pericolosità dell'acqua (WGK)

WGK 3

Punto d’infiammabilità (°F)

188.6 °F

Punto d’infiammabilità (°C)

87 °C

Dispositivi di protezione individuale

Eyeshields, Gloves, type ABEK (EN14387) respirator filter


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Methylation of single-stranded DNA in vitro introduces new restriction endonuclease cleavage sites.
B Gronenborn et al.
Nature, 272(5651), 375-377 (1978-03-23)
Regulation of the Escherichia coli lac operon expressed in human cells.
Bioard, D.S. et al.
Biochimica et Biophysica Acta, 113, 68-74 (1992)
Ausubel, F.M., et al.
Current Protocols in Molecular Biology, 1-1 (1995)
Characterization by in vitro complementation of a peptide corresponding to an operator-proximal segment of the beta-galactosidase structural gene of Escherichia coli.
A Ullmann et al.
Journal of molecular biology, 24(2), 339-343 (1967-03-14)
D S Biard et al.
Biochimica et biophysica acta, 1130(1), 68-74 (1992-02-28)
We have investigated the use of various Epstein-Barr virus (EBV)-based vectors bearing the two components of the Escherichia coli lac operator-repressor (lacO, lacI) complex. Our aim was to develop a model system of gene expression by looking at the transcription

Protocolli

Blue/white color selection streamlines vector insert differentiation, enhancing cloning efficiency for molecular biologists.

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