LIG1

DNA Ligation Kit

• UNSPSC Code: 12352204
• NACRES: NA.53

Properties

• grade: for molecular biology
• usage: kit sufficient for 150 ligation reactions
• shipped in: wet ice
• storage temp.: −20°C

Description

General description

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.

Application

Suitable for:

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

Components

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)

Principle

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

• Pictograms: GHS07
• Signal Word: Warning
• Hazard Statements: H319
• Precautionary Statements: P264 - P280 - P305 + P351 + P338 - P337 + P313
• Hazard Classifications: Eye Irrit. 2
• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable