Forestry and Forest Products Research Institute, Tsukuba, Ibaraki 305-8687, Japan.
Mol Ecol Resour. 2009 Jan;9(1):41-5. doi: 10.1111/j.1755-0998.2008.02184.x. Epub 2008 Oct 22.
We developed an efficient method for high-throughput extraction of high-quality DNA from various fungi. In this method, fungal mycelia were cultured and harvested on the surfaces of membranes on media plates. We degraded cell walls using a lytic enzyme (Yatalase). Purification was performed on 96-well glass fibre filter plates. DNA was successfully extracted from various fungi provided (102 genus 132 species) at high yields and quality, and proved suitable for storage, polymerase chain reaction amplification and restriction enzyme digestion. The method described is rapid, inexpensive and automation friendly. This enables the simultaneous extraction of large numbers of samples, significantly improving the potential throughput in genomics, particularly in diagnostic and population studies.
我们开发了一种从各种真菌中高通量提取高质量 DNA 的有效方法。在该方法中,真菌菌丝在培养基平板上的膜表面上进行培养和收获。我们使用溶菌酶(Yatalase)降解细胞壁。在 96 孔玻璃纤维滤板上进行纯化。成功地从各种真菌中提取了高产量和高质量的 DNA,并且证明适用于储存、聚合酶链反应扩增和限制酶消化。所描述的方法快速、廉价且易于自动化。这使得能够同时提取大量样品,极大地提高了基因组学中的潜在通量,特别是在诊断和种群研究中。