T4 DNA Polymerase, LIC-qualified - Novagen (C15-1307-834)

General description

T4 DNA polymerase is a potent enzymatic tool used in the process of ligation-independent cloning (LIC), a technique that allows to insert DNA fragments into plasmid vectors without the need for traditional restriction enzyme digestion and ligation steps. The 3′ exonuclease activity of T4 DNA polymerase is used in the LIC process to create single-stranded overhangs on the DNA fragment, which can then anneal with complementary overhangs on the vector, allowing the fragment to be inserted. In the LIC process, T4 DNA polymerase, in conjunction with complementary primers designed to lack one of dATP, dTTP, dCTP, or dGTP for a sequence of 12–15 bp, is used to generate single-stranded complementary overhangs of a fixed length. This annealing technique can then be followed by transformation directly into E. coli, which repairs the nicked DNA backbone, resulting in the successful insertion of the DNA fragment into the vector

Application

T4 DNA Polymerase, LIC-qualified – Novagen has been used in the ligation-independent cloning (LIC) reaction to generate single-stranded 5′ termini overhangs on the DNA insert and destination plasmid vector.

Components

•250 UT4 DNA Polymerase, LIC-qualified

•500 µl10X T4DNA Polymerase Buffer

Warning

Toxicity: Standard Handling (A)

Legal Information

NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany

Quality Level: 100. form: liquid. manufacturer/tradename: Novagen^®. . storage condition: OK to freeze, avoid repeated freeze/thaw cycles. technique(s): molecular cloning: suitable. storage temp.: −. 20°C. Storage Class Code: 10 - Combustible liquids. WGK: WGK 2.