Lamour Kurt, Finley Ledare
Department of Entomology and Plant Pathology, The University of Tennessee, Institute of Agriculture, Knoxville, Tennessee, USA,
Mycologia. 2006 May-Jun;98(3):514-7. doi: 10.3852/mycologia.98.3.514.
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.
我们提出了一种从大量疫霉菌分离物中回收高分子量基因组DNA的策略。其中包括在24孔可重复使用的深孔板中生成菌丝体团的步骤、高效冻干和破坏菌丝体以及使用96孔玻璃纤维滤板提取基因组DNA。所得DNA始终为高分子量,适用于需要高质量DNA的应用,如AFLP分析和TILLING。单个操作人员一天内可轻松处理384个分离物的菌丝体制备和/或DNA提取,该方法可能对其他可在液体培养基中生长的真菌或类真菌生物有用。