Mentor/s
Ashley Stoehr
Participation Type
Poster
Abstract
Meiofauna are a diverse group of benthic invertebrates that range between 0.04 mm and 1mm in size. They represent 24 of the 35 animal phyla and are ideal for biomonitoring. Their fast population turnover means that community composition can indicate anthropogenic influence and environmental health. The long-term goal of this project is to determine if visual identification of meiofauna at the level of phyla is sufficient to investigate community changes along a disturbed shoreline (Seaside Park; Bridgeport, CT); or if it is necessary to identify lower taxonomic levels via DNA metabarcoding. The latter would allow for the identification of taxa at genus or species levels based on a short variable region in the DNA. At present, this study aimed to optimize the DNA extraction protocol from samples containing meiofauna and sediment using the Qiagen DNeasy PowerSoil Pro Kit. Specific areas investigated included methods of sample storage (-80C vs. ethanol) and cell lysis (vortex vs. Mini BeadBeater-8). The combined use of frozen samples and the Mini BeadBeater-8 yielded more concentrated and purer samples of DNA. However, PCR was still necessary to visualize any DNA fragments following gel electrophoresis. Future work will include continued optimization of protocols, sequencing, and analysis.
College and Major available
Biology, Molecular & Cellular Biology BS
Location
Digital Commons & West Campus 2nd Floor University Commons
Start Day/Time
4-28-2023 12:00 PM
End Day/Time
4-28-2023 2:00 PM
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 License.
Optimization of a DNA Extraction Protocol for Meiofaunal Identification
Digital Commons & West Campus 2nd Floor University Commons
Meiofauna are a diverse group of benthic invertebrates that range between 0.04 mm and 1mm in size. They represent 24 of the 35 animal phyla and are ideal for biomonitoring. Their fast population turnover means that community composition can indicate anthropogenic influence and environmental health. The long-term goal of this project is to determine if visual identification of meiofauna at the level of phyla is sufficient to investigate community changes along a disturbed shoreline (Seaside Park; Bridgeport, CT); or if it is necessary to identify lower taxonomic levels via DNA metabarcoding. The latter would allow for the identification of taxa at genus or species levels based on a short variable region in the DNA. At present, this study aimed to optimize the DNA extraction protocol from samples containing meiofauna and sediment using the Qiagen DNeasy PowerSoil Pro Kit. Specific areas investigated included methods of sample storage (-80C vs. ethanol) and cell lysis (vortex vs. Mini BeadBeater-8). The combined use of frozen samples and the Mini BeadBeater-8 yielded more concentrated and purer samples of DNA. However, PCR was still necessary to visualize any DNA fragments following gel electrophoresis. Future work will include continued optimization of protocols, sequencing, and analysis.
Students' Information
Lilly Bove: molecular and cellular biology major, chemistry minor, graduating 2023
Joanna Coreno: molecular and cellular biology major, chemistry minor, psychology minor, honors minor, graduating 2023