Isolation of megabase-size DNA from sorghum and applications for physical mapping and bacterial and yeast artificial chromosome library construction

Authors

Sung-Sick Woo, Vipin K. Rastogi, Hong-Bin Zhang, Andrew H. Paterson, Keith F. Schertz and Rod A. Wing

Plant Molecular Biology Reporter

Volume 13, Number 1, 82-94, DOI: 10.1007/BF02668396

Abstract

A method was developed for the isolation of megabase-size DNA from Sorghum bicolor. Sorghum protoplasts were isolated from young leaf tissue, embedded in an agarose matrix as microbeads or plugs, followed by cell lysis and protein degradation. The DNA prepared by this method was larger than 1 Mb in size and readily digestible with restriction enzymes. The DNA was shown to be suitable for physical mapping, and was successfully used for the construction of BAC and YAC libraries.