Honors Theses

Date of Award

2016

Document Type

Undergraduate Thesis

Department

Biology

First Advisor

Erik Hom

Relational Format

Dissertation/Thesis

Abstract

Extracting high-quality RNA is critical for downstream applications such a qRT-PCR and RNA-Seq based transcriptomics. Single-stranded RNA is readily susceptible to degradation by environmental RNases, necessitating rapid and sterile homogenization techniques; freezing at -80°C, lysis buffer addition, and short bursts of beadbeating have been shown to increase yields and quality of extracted RNA. Utilizing fungal-algal tissues from the model symbiosis between the filamentous fungi Aspergillus nidulans and the green alga Chlamydomonas reinhardtii, three different commercial homogenizers and six RNA extractions kits were assessed and an optimized extraction protocol for total RNA from fungal-algal tissues was determined. Using the protocol designed in this study, the relationship between Chlamydomonas reinhardtii and the budding yeast Saccharomyces cerevisiae when grown under circadian-based light conditions prior to co-culturing will be assessed using RNA-Seq to determine changes gene expression patterns in response to co-culturing under a new light condition to look for synchrony. A phenotypic study of this system was performed and while no significant differences in biomass were observed, it is believed that with the fungus Neurospora crassa, a model organism for studying circadian rhythms, a light regimen-dependent phenotypic resonse will be observed.

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