An inexpensive high-throughput method to extract high yields of good quality DNA from fungi
An inexpensive high-throughput method to extract high yields of good quality DNA from fungi
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DOI：
10.1111/j.1755-0998.2008.02184.x
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发表时间：
2009-01-01
影响因子：
7.7
通讯作者：
Kubono, T.
中科院分区：
文献类型：
--
作者：
Kikuchi, T.;Karim, N.;Kubono, T.
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，真菌菌丝体在培养基平板上的膜表面上培养并收获。我们使用裂解酶（Yatalase）降解细胞壁。在96孔玻璃纤维滤板上进行纯化。成功地从提供的各种真菌（102属132种）中提取DNA，产量和质量都很高，并证明适合于储存、聚合酶链反应扩增和限制性内切酶消化。所描述的方法是快速，廉价和自动化友好。这使得能够同时提取大量样品，显著提高基因组学的潜在通量，特别是在诊断和人口研究中。
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.