RDLIG-RO

Rapid DNA Dephos & Ligation Kit

sufficient for 40 reactions (04898117001), sufficient for 160 reactions (04898125001), optimum reaction temp. 37 °C

• Synonym(s): cloning of dna fragments, dna ligase, ligases
• Enzyme Commission number: 3.1.3.1 (BRENDA, IUBMB)

Properties

• usage: sufficient for 160 reactions (04898125001); sufficient for 40 reactions (04898117001)
• Quality Level: 100
• manufacturer/tradename: Roche
• parameter: 37 °C optimum reaction temp.
• storage temp.: −20°C

Description

General description

The Rapid DNA Dephos & Ligation Kit enables fast and efficient dephosphorylation and ligation of sticky- or blunt-end DNA fragments.

Specificity

Specificity: rAPid Alkaline Phosphatase
Alkaline Phosphatase catalyzes the hydrolysis of numerous phosphate esters, such as esters of primary and secondary alcohols, saccharides, cyclic alcohols, phenols and amines. Phosphodiesters do not react. The enzyme hydrolyzes inorganic pyrophosphate. The kinetic properties of the enzyme depend on many factors, such as purity of enzyme, concentration of enzyme in the assay, buffer, pH etc.
Heat inactivation: rApid Alkaline Phosphatase:
Two minutes at 75 °C.
T4 DNA Ligase:
10 minutes at 65 °C.

Application

The Rapid DNA Dephos & Ligation Kit minimizes the time needed to perform diverse applications, including:

• Cloning of fragments into either plasmid or phage vectors[1]
• Linker ligation
• Recircularization of linear vector DNA
• Generation of libraries

Packaging

1 kit containing 5 components

Unit Definition

Unit Definition: rAPid Alkaline Phosphatase
One unit of rAPid Alkaline Phosphatase is the enzyme activity which hydrolyzes one μmol of 4-nitrophenyl phosphate in one minute at +37 °C under assay conditions.

Volume Activity: rApid Alkaline Phosphatase: 1 U/μl
T4 DNA Ligase: 5 U/μl

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

• rAPid Alkaline Phosphatase Buffer 10x concentrated
• rAPid Alkaline Phosphatase 1 U/μl
• DNA Dilution Buffer 5x concentrated
• T4 DNA Ligation Buffer 2x concentrated
• T4 DNA Ligase 5 U/μl

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash