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Company: Grainger
Catalog#: 4YR61
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Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions
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2020-06-05
[Abstract]  We present a safe and low-cost method suitable for DNA extraction from mycelium and tree tissue samples. After sample preparation, the extraction takes about 60 min. Method performance was tested by extracting DNA from various tree tissue samples and from mycelium grown on solid and liquid media. DNA was extracted from juvenile and mature host material (Picea abies, Populus trichocarpa, Pseudotsuga menziesii) infected with different pathogens (Heterobasidion annosum, Heterobasidion parviporum, Leptographium wagenerii, Sphaerulina musiva). Additionally, DNA was extracted from pure cultures of the pathogens and several endophytic fungi. PCR success rate was 100% for young poplar material and fungal samples, and 48-72% for ... [摘要]  [摘要 ] 我们提出了一种安全,低成本的方法,适用于从菌丝体和树木组织样品中提取DNA。样品制备后,提取过程大约需要60分钟。通过从各种树木组织样品和菌丝体中提取DNA来测试方法性能DNA从少年提取和成熟主体材料(云杉冷杉,胡杨毛果,黄杉花旗松感染不同病原体()Heterobasidion annosum ,Heterobasidion parviporum ,Leptographium wagenerii ,Sphaerulina musiva )。额外的,DNA从提取纯的病原体和几种内生真菌培养物。年轻杨树材料和真菌样品的PCR成功率是100%,针叶树和成熟阔叶植物样品的PCR成功率是48-72%。我们建议使用10-50 mg的新鲜样品最好结果。该方法为研究树-真菌相互作用提供了一种安全且低成本的DNA提取方法,并且是潜在的提纯方法。兴的目的。

[背景 ] DNA提取是植物-微生物相互作用研究和植物疾病诊断purposes.The丰植物真菌相互作用的中央技术由高数量从植物样品中分离的培养的菌株的反射(阿诺德等人,2001;希金斯等人,2007; Terhonen 等人,2011)。使用商业试剂盒处理大量样品进行常规PCR反应的成本可能很高,此外,对2-巯基乙醇,氯仿或苯酚等有害化学品的需求也可能增加。限制协议在教学和培训中的适用性。

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