2021 Summer Undergraduate Research Program (SURP) Symposium

Location

Ballroom, Maucker Student Union, University of Northern Iowa

Presentation Type

Open Access Poster Presentation

Document Type

poster

Keywords

Fiddler crabs--Genetics;

Abstract

Animal mitochondrial DNA can serve as an important model in understanding human mitochondrial DNA and the mutations that may be associated in causing human mitochondrial diseases, which have not been widely studied. In order for animal models to be meaningful, the variation among genomes within evolutionary groups must first be understood. The fiddler crab family (Ocypodidae) is a widespread animal family with an increasing number of described mitogenomes. One of which being a yet unexamined taxon, Leptuca spinicarpa, the spiny-wristed fiddler crab that lives in North America along the shores of the Gulf of Mexico where it is periodically submerged by brackish tides. Previously, the mitochondrial genome of L. spinicarpa was partially sequenced using NGS (Next-Generation Sequencing) Illumina Sequencing, but three regions of difficulty, or gaps, remained. Using the mitogenomic sequence of Minuca minax (LeConte 1855) as a reference, we used polymerase chain reactions (PCR) to create copies of the missing DNA sequences in L. spinicarpa’s mitogenome. Custom designed primers, or the short sequences of DNA that jumpstart the process of replication, were created through careful examination of L. spinicarpas nucleotide bases. Possible primers included sequences that were around 20 base pairs in length, starting with the GC nucleotides. Primers were located about 100-500 base pairs away from the missing sequence, or gap. PCR was performed with several different combinations of forward and reverse primers. Results were then quantified using gel electrophoresis. Once the gel confirmed that the PCR successfully amplified the DNA sequence in question, Sanger sequencing was used for these relatively short regions (ca. 1000 nucleotides). As a result, two of the three gaps are now fully sequenced. These results suggest that L. spinicarpa has a similar gene arrangement to M. minax, which will be helpful in reassessment of the phylogenetic relationships within the fiddler crab family.

Start Date

30-7-2021 11:30 AM

End Date

30-7-2021 1:15 PM

Event Host

Summer Undergraduate Research Program, University of Northern Iowa

Faculty Advisor

James DeMastes

Department

Department of Biology

Creative Commons License

Creative Commons Attribution-NonCommercial 4.0 International License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License

File Format

application/pdf

Available for download on Wednesday, October 19, 2022

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Jul 30th, 11:30 AM Jul 30th, 1:15 PM

The Mitochondrial Genome of the Spiny-Wristed Fiddler Crab, Leptuca Spinicarpa Rathbun 1900

Ballroom, Maucker Student Union, University of Northern Iowa

Animal mitochondrial DNA can serve as an important model in understanding human mitochondrial DNA and the mutations that may be associated in causing human mitochondrial diseases, which have not been widely studied. In order for animal models to be meaningful, the variation among genomes within evolutionary groups must first be understood. The fiddler crab family (Ocypodidae) is a widespread animal family with an increasing number of described mitogenomes. One of which being a yet unexamined taxon, Leptuca spinicarpa, the spiny-wristed fiddler crab that lives in North America along the shores of the Gulf of Mexico where it is periodically submerged by brackish tides. Previously, the mitochondrial genome of L. spinicarpa was partially sequenced using NGS (Next-Generation Sequencing) Illumina Sequencing, but three regions of difficulty, or gaps, remained. Using the mitogenomic sequence of Minuca minax (LeConte 1855) as a reference, we used polymerase chain reactions (PCR) to create copies of the missing DNA sequences in L. spinicarpa’s mitogenome. Custom designed primers, or the short sequences of DNA that jumpstart the process of replication, were created through careful examination of L. spinicarpas nucleotide bases. Possible primers included sequences that were around 20 base pairs in length, starting with the GC nucleotides. Primers were located about 100-500 base pairs away from the missing sequence, or gap. PCR was performed with several different combinations of forward and reverse primers. Results were then quantified using gel electrophoresis. Once the gel confirmed that the PCR successfully amplified the DNA sequence in question, Sanger sequencing was used for these relatively short regions (ca. 1000 nucleotides). As a result, two of the three gaps are now fully sequenced. These results suggest that L. spinicarpa has a similar gene arrangement to M. minax, which will be helpful in reassessment of the phylogenetic relationships within the fiddler crab family.