A strategy for recovering high quality genomic DNA from a large number of Phytophthora isolates.
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.
Lamour
Kurt
K
Department of Entomology and Plant Pathology, The University of Tennessee, Institute of Agriculture, Knoxville, Tennessee, USA, klamour@utk.edu
Finley
Ledare
L
D003470:Culture Media; D004247:DNA / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D005821:Genetic Techniques; D016678:Genome / Q000235:genetics; D009154:Mutation; D010838:Phytophthora / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D012150:Polymorphism, Restriction Fragment Length
D003470:Culture Media; D004247:DNA