E-Book Overview
Erich Grotewold has assembled a team of leading plant scientists to describe in detail the most commonly used methods for investigating plant gene function in a wide variety of plants, during plant pathogen interactions, and even in algae. These readily reproducible protocols include computational, molecular, and genetic methodologies designed for both general and specific problems. Here the reader will learn about powerful computational and statistical tools to help predict gene function either on the basis of comparative genomics, or from the analysis of complex genome sequences. Numerous loss-of-function and gain-of-function techniques for discovering gene function are presented in step-by-step detail.
E-Book Content
Methods in Molecular Biology
TM
VOLUME 236
Plant Functional Genomics Edited by
Erich Grotewold
Plant BAC Library Construction
3
1 An Improved Method for Plant BAC Library Construction Meizhong Luo and Rod A. Wing
Summary Large genomic DNA insert-containing libraries are required as critical tools for physical mapping, positional cloning, and genome sequencing of complex genomes. The bacterial artificial chromosome (BAC) cloning system has become a dominant system over others to clone large genomic DNA inserts. As the costs of positional cloning, physical mapping, and genome sequencing continuously decrease, there is an increasing demand for high-quality deepcoverage large insert BAC libraries. In our laboratory, we have constructed many high-quality deep-coverage large insert BAC libraries including arabidopsis, manocot and dicot crop plants, and plant pathogens. Here, we present the protocol used in our laboratory to construct BAC libraries.
Key Words BAC, library, method, pCUGIBAC1, plant
1. Introduction Large genomic DNA insert-containing libraries are essential for