Genomic DNA Variation Between Orb, Sheet Web, and Ground Dwelling Spiders

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Genomic DNA Variation Between Orb, Sheet Web, and
Ground Dwelling Spiders

• Bellarmine University
• Department of Biology
• Louisville, KY
• Stephanie Sermon

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Introduction

• There are 35,000 species of named spiders
  worldwide, but the majority of spiders have not
  yet been identified.
• Currently spiders are classified based on
  physical characteristics.
• The goal of this research is to inspect
  characteristics and relationship between
  different species of spiders at the genomic level.

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Procedures for Research

• Collection of spiders
• DNA extraction
• Spectrophotometer
• RAPD-PCR
• Gel Electrophoresis

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Eight Species of Spiders were Observed in this
Research

• Four ground dwellers.
• Two orb weavers.
• Two sheet web builders.

• Lycosa rabida, Agelena labyrinthica, Kukulcania
  hibernalis, and Tidarren sisyphoides.
• Araneus angulatus and Neriene radiata.
• Frontinella pyramitella and Florinda coccina.

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Ground Dwellers

• Family Lycosidae.
• They have 8 eyes of unequal size in arranged in 3
  rows.
• The abdomen and thoraxes are usually long.
• They are usually brown with dark spots.

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Orb Weavers

• Family Araneidae.
• They are usually 1/16 1 1/8 long.
• Eight eyes in two horizontal rows of four each.
• Spin big, usually circular webs.

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Sheet Web Builder

• Family Linyphildae
• Long slender legs with short bristles.
• Eight eyes in 2 parallel rows.
• 1/16 3/8 long.
• Flat sheet like or dome shaped webs.

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Collection of Spiders

• The orb weavers were collected from parking
  garage of Caesars Indiana.
• The sheet web builders were collected from the
  grass and bushes outside of Pasteur hall.
• The ground dwellers were captured from various
  houses.

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DNA Extraction

• Genomic DNA was extracted using the Promega
  Wizard Genomic DNA Purification kit.
• This process involved using a nuclei lysis
  solution, Rnase solution, a protein precipitation
  and a DNA rehydration solution.

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Spectrophotometer

• Spectrophotometric absorbances were measured at
  280nm, 260nm, and 234nm.
• Absorbance ratios had to lie between 1.6-2.0 for
  260/280. This is the DNARNA ratio.
• TE buffer was used as the blank sample.

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RAPD-PCR

• Random amplified polymorphic DNA analysis.
• RAPD analysis enables quick detections of
  polymorphisms at a number of different loci while
  using very small quantities of genomic DNA.
• Primer sequence (5 dGTAGACCCGT-3)

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Gel Electrophoresis

• A mini-gel set up was used for eight samples with
  a 100bp molecular weight ladder on each side.
• Electrophoresis was carried out in 1XTBE, and
  electrophoresed at 175V for 35 minutes.

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Ethidium Bromide

• Ethidium Bromide is the best stain for observing
  genomic DNA.
• Although, it wasnt possible to get a measurable
  picture by digital camera.

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Gel Analysis

• Photos were then taken by Polaroid.
• The distance that the band moved was measured in
  cm and recorded on logarithmic graphing paper to
  determine the molecular weight.

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Results

• A Orb weavers vs. Ground dwellers
• B Orb weavers vs. Sheet web builders
• C Ground Dwellers
• D Ground Dwellers vs. Sheet web builders

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Measurements of Bands
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Comparing all 8 Species
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Results

• Same types of spider different species
• Agelena labyrinthica and Lycosa rabida
• Kykulcania hibernalis and Agelena labyrinthica
• Florinda coccina and Frontinella pyramitella

• Different types of spiders closely related
• Kukulcania hibernalis and Florinda coccina
• Agelena labyrinthica and Araneus angulatus

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Conclusion

• Ground Dwellers were more genetically related
  to both sheet web builders and orb weavers, than
  orb weavers to sheet web builders.
• RAPD-PCR is a useful process in detecting
  polymorphisms between many different species of
  spiders.

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References

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