Insert selectable markers into microorganisms, in vivo . Strategy Prepare Ordered Deletion Templates for Sequencing, Find Precise Gene Coding Sequences, or Map Epitopes. Transposon Tools provide a simple, quick and reliable method to generate random, unidirectional deletions in plasmid and cosmid clones. The process for producing and identifying deletion clones is summarized below. Reaction conditions have been optimized to maximize deletion events in a 2 hour to overnight incubation.
Intramolecular transposition yields two types of products.
In the second case transposition results in an inversion of one portion of the cloned insert. Since the nature and the site of the intramolecular reaction is purely random, the result is the generation of a population of independent clones, each containing either a unidirectional deletion or inversion. Since all of the inversion clones are the same size, deletion clones can be easily distinguished from inversion clones based on size analysis by agarose gel electrophoresis. 8 Applications of the In Vitro Deletion Strategy Identifying the precise coding sequence of cloned genes. DNA sequencing template using sequencing primers provided in the Kits.