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About This Item
NACRES:
NA.54
UNSPSC Code:
12352202
Quality Segment
form
liquid
manufacturer/tradename
Novagen®
storage condition
OK to freeze, avoid repeated freeze/thaw cycles
technique(s)
molecular cloning: suitable
storage temp.
−20°C
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
Other Notes
Legal Information
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)
Storage Class
10 - Combustible liquids
WGK
WGK 1
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