A highly efficient method for concentrating DNA from river water by combined coagulation and foam separation
A highly efficient method for concentrating DNA from river water by combined coagulation and foam separation
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DOI：
10.1080/01496395.2019.1565774
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发表时间：
2019-01
影响因子：
2.8
通讯作者：
Yoshihiro Suzuki;Yuki Imafuku;Masateru Nishiyama;Kotaro Teranishi;Atsushi Jikumaru;K. Nukazawa;
中科院分区：
文献类型：
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作者：
Yoshihiro Suzuki;Yuki Imafuku;Masateru Nishiyama;Kotaro Teranishi;Atsushi Jikumaru;K. Nukazawa;
摘要 使用氯化铁和牛奶酪蛋白通过混凝和泡沫分离，在 7 分钟内从 500 mL 加有细菌的水中收集到泡沫中的大肠杆菌和肠球菌。使用 DNeasy PowerWater® 试剂盒从泡沫中的 100 µL 提取物中分离出这些细菌 DNA，回收率超过 87.5%。为了用来自 3 条天然河流的水测试该方法，将 0.67–2.70 µg DNA 浓缩在 1,000 mL 河水中的 100 µL 提取物中。当DNA提取物进行16S rRNA基因测序分析时，可以获得细菌菌群的信息。
ABSTRACT Both Escherichia coli and Enterococci were collected in foam within 7 min from 500 mL of bacteria-spiked water by coagulation and foam separation using ferric chloride and milk casein. These bacterial DNA isolated in the 100 µL of extract from the foam more than 87.5% recovery using the DNeasy PowerWater® Kit. To test this method with water from three natural rivers, 0.67–2.70 µg of DNA were concentrated in 100 µL of extract from 1,000 mL of river water. When the DNA extract was subjected to 16S rRNA gene sequencing analysis, information on the bacterial flora could be obtained.