Cloning involves the insertion of a donor DNA fragment into a suitable vector via a process known as ligation. Ligation is performed by DNA ligase, which forms covalent phosphodiester bonds between the 3′-hydroxyl and 5′-phosphate groups of terminal nucleotides of the donor insert and vector DNA. Cloned DNA fragments are typically used for sequence characterization or functional characterization of genes, techniques that require high-quality DNA inserts with no spurious mutations.
In the modern age of molecular biology, PCR isolation of target gene products has become standard and requires polymerase enzymes that yield high-quality products for downstream analysis. Kapa Biosystems has evolved a proofreading polymerase that exhibits exceptional fidelity (100X lower error rate than Taq polymerase), sensitivity and robustness, making it ideally suited for cloning applications sensitive to spurious mutations.
For Research Use Only. Not for use in diagnostic procedures.