LIG1

DNA Ligation Kit

• NACRES: NA.53

Properties

• grado: for molecular biology
• Nivel de calidad: 200
• uso: 1 kit sufficient for 150 ligation reactions
• enviado en: wet ice
• temp. de almacenamiento: −20°C

Description

Descripción general

Sigma′s DNA Ligation Kit contains all of the reagents necessary to perform DNA ligation reactions. Sticky-end ligations are more efficient than blunt-end ligations; these can be facilitated with the addition of PEG.

Aplicación

Suitable for:

• Joining fragments of DNA into a cloning vector
• Mutagenesis
• Gene anyalysis and structure-function relationships

Componentes

Sufficient for 150 reactions:

• 300uL 10X Ligation Buffer (D2176) in 250 mM Tris-HCl (pH 7.8) 100 mM MgCl2, and 10 mM dithiothreitol
• 3 x 100 units T4 DNA Ligase (D2886) in 50% glycerol with 10 mM Tris-HCl (pH 7.5) 50 mM KCl, and 1 mM dithiothreitol
• 3 x 100 uL 10 mM ATP ( A3702)
• 50 uL Control pBR322 DNA, HAE III Digest (D9430) 0.5 ug/ul in 10 mM Tris-HCl (pH 8.0), and 1 mM EDTA
• 1.5 mL 24% (w/v) PEG Solution, (P 2454)
• 1.5 mL Molecular Biology Grade Water (W4502)

Principio

One of the most important steps in the cloning process is the ligation of linear DNA into a cloning vector. DNA ligations are performed by incubating DNA fragments with appropriately linearized cloning vectors in the presence of buffer, ATP, and DNA ligase. Many parameters affect ligations such as the relative ratio of insert to vector, the quality and type of the DNA ends, the temperature of ligation and the concentration of DNA.

Safety Information

• pictogramas: GHS07
• Palabra de señalización: Warning
• Frases de peligro: H319
• Consejos de prudencia: P264 - P280 - P305 + P351 + P338 - P337 + P313
• Clasificaciones de peligro: Eye Irrit. 2
• Código de clase de almacenamiento: 10 - Combustible liquids
• Punto de inflamabilidad F: Not applicable
• Punto de inflamabilidad C: Not applicable