Speakers:
Jim Pease, EPICENTRE

Workshop Information: Recent Advances that Simplify and Speed Up Sequencing of Plasmid, Cosmid and BAC Clones

Sequencing of large plasmid, cosmid and Bacterial Artificial Chromosome (BAC) clones can be an arduous project that is hampered by inefficiencies. Subcloning of large cloned inserts followed by shotgun sequencing is very laborious and labor-intensive. Sequencing large clones by primer walking can be costly and time consuming. A faster and more efficient approach to sequencing large plasmid and cosmid clones is by transposon-mediated sequencing (TMS). TSM, in combination with high yield BAC clone purification methods (the PSIClone Kit) even facilitates direct sequencing of large, low copy BAC clones without the need for subcloning.

In this workshop we describe a new transposon-based strategy (EZ::TN Transposon Tools) to facilitate sequencing of large clones faster and easier than by subcloning or primer walking. The first half of the presentation will introduce the EZ::TN Transposon Tools and their use in efficiently and rapidly generating a complete population of DNA sequencing template from plasmid, cosmid and BAC clones. The second part of the workshop will focus on purification of and direct transposon-mediated sequencing of BAC clones. Laboratory data and practical tips for direct sequencing large plasmid, cosmid and BAC clones will be presented.

This page last updated Sunday, 03-Dec-2000 13:59:26 EST