2025 Summer Undergraduate Research Program (SURP) symposium

Location

Dr. Ken Budke Family Auditorium, Schindler Education Center, University of Nothern Iowa

Presentation Type

Open Access Poster Presentation

Document Type

poster

Abstract

16s metabarcoding uses the ubiquitous prokaryotic 16s mRNA sequence as a “DNA fingerprint” to identify bacteria and archaea by their unique 16s sequence. Caves offer a unique environment to study microbial life that survives under extreme nutrient limitation. To this extent, 16s metabarcoding allows us to identify and study the microbial composition of these environments, but there are hurdles that must be overcome to obtain enough DNA for effective and reliable 16s sequencing.

Wind Cave National Park serves as a relatively accessible cave microbiome to study. It is a limestone karst cave, meaning it is almost entirely made up of the minerals calcite and chert. Minerals, especially calcite, can inhibit DNA extraction because of positive charges in the mineral structure trapping negatively charged DNA on its surface [1-3, 5]. Our lab has previously worked on developing a DNA extraction protocol suited for high DNA yield from environmental samples, including many experiments with Wind Cave samples. It is important to maximize the amount of DNA that can be extracted from the Wind Cave samples because it is an extremely low biomass environment and sufficient DNA for sequencing can be hard to get.

I’ve been developing and testing a new DNA extraction method optimized for mineralogic samples, named the Mineral DNA extraction protocol.

Start Date

1-8-2025 11:00 AM

End Date

1-8-2025 1:30 PM

Event Host

Summer Undergraduate Research Program, University of Northern Iowa

Faculty Advisor

Marek Sliwinski

Department

Department of Biology

File Format

application/pdf

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Aug 1st, 11:00 AM Aug 1st, 1:30 PM

Developing a DNA Extraction Method for Mineral Abundant Limestone Cave Samples

Dr. Ken Budke Family Auditorium, Schindler Education Center, University of Nothern Iowa

16s metabarcoding uses the ubiquitous prokaryotic 16s mRNA sequence as a “DNA fingerprint” to identify bacteria and archaea by their unique 16s sequence. Caves offer a unique environment to study microbial life that survives under extreme nutrient limitation. To this extent, 16s metabarcoding allows us to identify and study the microbial composition of these environments, but there are hurdles that must be overcome to obtain enough DNA for effective and reliable 16s sequencing.

Wind Cave National Park serves as a relatively accessible cave microbiome to study. It is a limestone karst cave, meaning it is almost entirely made up of the minerals calcite and chert. Minerals, especially calcite, can inhibit DNA extraction because of positive charges in the mineral structure trapping negatively charged DNA on its surface [1-3, 5]. Our lab has previously worked on developing a DNA extraction protocol suited for high DNA yield from environmental samples, including many experiments with Wind Cave samples. It is important to maximize the amount of DNA that can be extracted from the Wind Cave samples because it is an extremely low biomass environment and sufficient DNA for sequencing can be hard to get.

I’ve been developing and testing a new DNA extraction method optimized for mineralogic samples, named the Mineral DNA extraction protocol.