DNA Cloning & Ligation
DNA cloning has become an established method to insert DNA fragments from any source into a cloning vector using restriction enzymes and DNA ligases. The resulting recombinant DNA molecules are then introduced mostly into bacterial cells, but also viruses and sporadically eukaryotic cells (via transfection and transformation). Inside the host cells the DNA molecules are duplicated. There is therefore an almost unlimited amount of DNA. These DNA segments are then tested for accuracy, and used in other processes, for example in the production of recombinant proteins (for example, insulin, EGF).
DNA ligases catalyze the formation of a phosphodiester bond between the 3'-hydroxyl and 5'-phosphate termini of adjacent DNA residues. In living organisms, ligases play a crucial role in DNA repair and DNA replication. In vitro, the enzyme is used to connect double-stranded DNA fragments with blunt or cohesive ends, to form, for example, recombinant DNA plasmids or to connect DNA probes for SNP detection.
The T4 DNA ligase can connect both blunt ends as well as cohesive DNA ends in duplex DNA/RNA. It repairs single-strand nicks in duplex DNA, RNA and DNA/RNA hybrids.
For your successful cloning and ligation Genaxxon bioscience offers:
Chemically Competent Cells - T4 DNA ligase for ligation