A strategy for recovering high quality genomic DNA from a large number of Phytophthora isolates

Abstract

We present a strategy to recover high molecular weight genomic DNA from large numbers of isolates of Phytophthora. Included are steps for generating mycelial mass in 24-well reuseable deep well plates, efficient lyophilization and disruption of the mycelium and genomic DNA extraction with 96-well glass fiber filter plates. The resulting DNA is consistently high molecular weight and is suitable for applications that require high quality DNA such as AFLP analysis and TILLING. A single operator easily can manage mycelium preparation and/or DNA extraction from 384 isolates in a single day and this approach might be useful for other fungi or fungi-like organisms that can be grown in liquid media.

• AFLP
• oomycetes
• reverse genetics
• TILLING

This material is based on work supported by the National Science Foundation under Grant No. 0347624. Special thanks for the assistance of Dr Linda Cain, Sharyce Banks and Catherine Zama of Knoxville College and to Jason Mclean and Matt Smith of the University of Tennessee.