Bacterial artificial chromosome (BAC) libraries

The construction of bacterial artificial chromosome (BAC) libraries

SANGDUN CHOI 1,2 and ROD A. WING 2

1 Department of Soil and Crop Sciences, Texas A&M University, College Station, TX 77843, USA

2 Departments of Agronomy & Biological Sciences, Clemson University Genome Center, Clemson University, Clemson, SC 29634, USA

Introduction

Cloning of exogenous DNA into bacterial artificial chromosomes (BACs) provides a new approach to the analysis of the genomes of higher organisms [1]. BAC libraries containing large genomic DNA inserts are important tools for positional cloning, physical mapping and genome sequencing. A number of human and plant BAC libraries have been constructed (e.g., human: [2], Arabidopsis: [3], rice: [4], sorghum: [5]). Bacterial artificial chromosome vectors utilize the E. coli single-copy fertility plasmid and can maintain genomic DNA fragments up to 350 kb. Very little or no rearrangement of the inserts or chimerism have been observed [1, 5, 6, 7]. Other systems for the cloning of large DNA fragments have been developed. The development of yeast artificial chromosome vectors (YAC: [8]) permits cloning of fragme

Full Protocol Here

http://www.genome.clemson.edu/groups/bac/protocols/protocols2new.html