[Next-Generation Sequencing Analysis Case] Analysis of Live Bacterial Microbial Community Structure
Introducing a case that demonstrates the method using EMA can analyze live microbial communities! *Case study collection available.
We will introduce community structure analysis using the selective membrane permeable dye EMA (in-house data). For environmental samples dominated by E. coli, we prepared samples that were heat-treated and sterilized, as well as untreated samples, and performed EMA treatment. Subsequently, DNA extraction was conducted for each sample, followed by 16S rRNA gene amplicon analysis. As a result, in the community structure analysis, E. coli, which had become dominant, was hardly detected in the heat-treated samples due to its death. Additionally, in the related links, we also introduce community structure analysis using reverse transcription cDNA from RNA (in-house data), so please take a look. 【Workflow】 ■ Prepare environmental samples (heat-treated, untreated) ■ EMA treatment ■ Light irradiation ■ DNA extraction ■ Creation of adapter library via EMA-PCR *For more details, please refer to the PDF document or feel free to contact us.
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【Results】 - For samples that were not treated with EMA, there were no differences observed in both the DNA concentration of the library and the community structure. - On the other hand, for samples treated with EMA, the library concentration of heat-treated samples was low due to inhibition of PCR amplification. - In community structure analysis, the dominant E. coli was almost undetectable in heat-treated samples due to its death. - It is anticipated that DNA was inactivated by EMA, which inhibited PCR amplification. *For more details, please refer to the PDF document or feel free to contact us.*
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For more details, please refer to the PDF document or feel free to contact us.
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In recent years, advancements in molecular biology have led to remarkable progress in research such as the determination of the complete genome sequences of organisms and subsequent gene function analysis. Along with these technological advancements, molecular biological techniques such as nucleotide sequence analysis, real-time PCR, and DNA chips have become widely utilized in the field of food analysis. Since its establishment in 2001, Fasmak Co., Ltd. has been developing technologies for the "Japanese Standard Analysis Method" for genetically modified foods and food allergens in collaboration with related agencies of the Ministry of Agriculture, Forestry and Fisheries and the Ministry of Health, Labour and Welfare. The developed testing technologies are provided not only in Japan but also in the United States, China, and other countries. Additionally, Fasmak has been actively engaged in international standardization activities for "food inspection methods using molecular biological techniques" since its establishment, and its technical capabilities are internationally recognized. Furthermore, in partnership with Eurofins Scientific, one of the world's largest testing companies, Fasmak is advancing the introduction of the high technical capabilities possessed by the Eurofins Group. Moving forward, Fasmak will continue to provide world-class new testing technologies to everyone.