Tol2kit

The Tol2kit is a set of constructs that can be used with Invitrogen's 3-insert Multisite Gateway recombination-based cloning system to build expression constructs in a Tol2 transposon backbone (derived from constructs from the Kawakami lab). It has several advantages, including (1) ease of cloning large constructs in a Tol2 backbone; (2) reuse of existing components, including different promoters, fluorescent protein reporters, and IRES and 3' fusion tags; (3) the ability to transfer clones between labs in a standard format; and (4) the ability to add transgenesis markers such ascmlc2:egfp on the same plasmid backbone.

The Tol2kit is described in a paper in Developmental Dynamics; complete documentation including protocols and lists of constructs can be found at the Chien lab's Tol2kit documentation site. A related set of constructs based on 2-insert Multisite Gateway are described at the Lawson lab Gateway site.

This wiki page is intended for discussion and questions about the Tol2kit. The Chien lab will monitor posted comments and try to respond quickly. A public blog was originally used for this purpose; we are currently in the process of transferring questions and comments from the blog to this page.