Assessment of the DNA Quality in Mushroom Specimens: a Recovery of the Whole ITS Sequence from Fragmented DNA of the Type Specimen 1
Assessment of the DNA Quality in Mushroom Specimens: a Recovery of the Whole ITS Sequence from Fragmented DNA of the Type Specimen 1
复制标题
蘑菇样本 DNA 质量评估：从模式样本 1 的 DNA 片段中恢复整个 ITS 序列
DOI：
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
K. Uno
中科院分区：
文献类型：
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作者：
K. Hosaka;K. Uno
从20世纪50年代以来保存于国家自然科学博物馆植物标本室的蘑菇（木耳门、担子菌门、真菌门）4种类型标本中提取DNA，并进行PCR扩增。由于整个ITS区域（约650 bp）无法扩增，因此假定DNA高度降解至小于600 bp。为了评估降解程度，我们使用了基于微流体的平台。结果显示，最常见的片段长度小于230 bp。基于这一结果，我们设计了6条新的引物，可以特异性扩增约200- 400bp的Amanita imazekii产物。通过对8个序列的拼接，成功地重建了该物种的整个ITS区域，恢复的序列与参考序列的匹配率为100%。
DNA from four type specimens of mushrooms (Agaricales, Basidiomycota, Fungi) stored in the herbarium of National Museum of Nature and Science since 1950�s were extracted and PCR amplification was attempted. Because the whole ITS region (ca. 650 bp) could not be amplified, DNA were assumed to be highly degraded to less than 600 bp. To assess the degree of degradation, we used a microfluidics-based platform. This revealed that the most frequently pres- ent fragments were less than 230 bp long. Based on this result, we have designed six new primers to specifically amplify approximately 200-400 bp products of Amanita imazekii. A total of 8 con- tigs were assembled to successfully reconstruct the whole ITS region of the species, and the sequence recovered by this approach had a 100% match with the reference sequence.
DOI：
10.1073/pnas.1117018109
发表时间：
2012-04-17
影响因子：
11.1
作者：
Schoch, Conrad L.;Seifert, Keith A.;Schindel, David
通讯作者：
Schindel, David