Geneious Prime provides a tool for in silico simulation of TOPO-cloning.
TOPO vectors provide a fast and convenient method to reliably ligate PCR products (where 5' ends are not phosphorylated) in as little as 5 minutes at room temperature.
TOPO cloning utilises the Vacinnia DNA topoisomerase I enzyme, which functions both as a restriction enzyme and as a ligase.
In this tutorial we will learn how to clone Human Proinsulin, the precursor of insulin, into TOPO vectors using TA-, Blunt- and directional methods.
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Exercise 1: Cloning into TA Vector
Exercise 2: Cloning into ZeroBlunt Vector
Exercise 3: Directional Cloning
Exercise 4: Batch Cloning