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Multiplexed isotope in situ hybridization (iISH) to profile complex microbial assemblages


EMSL Project ID
60838

Abstract

The use of fluorescence based in situ hybridization (FISH) is an established method for identifying specific nucleic acids associated with microorganism identification (rRNA) and/or function (mRNA, DNA). However, certain methodological challenges (i.e., background, autofluorescence, spectral overlap) limit the number of targets/probes that can be analyzed within any one sample – be that a single cell or complex assemblage of individuals. We propose multiplexing numerous monoisotopic tags (iISH)
to target more DNA/RNA sequences at once. The advantage of this approach would be resolving more species or functions within complex microbial systems and improved understanding of spatial relationships between target organisms (or sub-cellular features).

Project Details

Start Date
2023-04-17
End Date
2023-11-10
Status
Closed

Team

Principal Investigator

Jeremy Bougoure
Institution
Environmental Molecular Sciences Laboratory

Co-Investigator(s)

Dehong Hu
Institution
Environmental Molecular Sciences Laboratory

Team Members

John Cliff
Institution
Environmental Molecular Sciences Laboratory