Development of high sencitive FISH for revealing functions of uncultured microorganisms

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K14807
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Aquatic life science
• Research Institution: Anan National College of Technology
• Principal Investigator: Kawakami Shuji 阿南工業高等専門学校, 創造技術工学科, 講師 (00610461)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 環境微生物 / シングルセル解析 / FISH法

Outline of Final Research Achievements

In this study, we introduced an improved in situ DNA-HCR protocol (quickHCR-FISH) with higher sensitivity, and the amplification time was accelerated to only 15 min by using a dextran sulfate and blocking reagent containing amplification buffer, compared with 2 h in the original protocol. Finally, quickHCR-FISH was applied to marine bacteria in seawater and sediment samples for checking applicability of environmental microorganisms. The detection rates of marine bacteria after quickHCR-FISH were almost the same as after CARD-FISH. This newly developed protocol can be an attractive alternative to CARD-FISH for the detection of microorganisms in environmental context.

Free Research Field

環境微生物学