Telomeres are nucleoprotein complexes at the ends of chromosomes and consist of non-coding, tandem repeats. They serve to protect chromosomal ends as well as ensure the complete transmission of genetic information from one generation to another. In a wide range of organisms, including humans, telomeres in germ cells are found to be significantly longer than those in somatic cells. We are interested in developing a simple model organism to study this phenomenon. Our lab uses the fungus Aspergillus nidulans due in part to its exceptionally short telomeres observed in vegetative cells. The telomeres in sexual cells have not been studied. We hypothesize that in this fungus, the sexual spores have longer telomeres than the vegetative cells, analogous to what is seen in other eukaryotes. The initial steps of testing this hypothesis involved opening the ascospores and subsequently isolating intact DNA from them. Though I encountered many failed experiments in attempting those initial steps, I was eventually able to develop a protocol for extracting intact DNA from ascospores to be used in a PCR assay to determine telomere length. No such extraction procedure was previously available. After obtaining DNA from ascospores for PCR, I used the anchored telomere PCR assay to determine the telomere length in the sexual spores of A. nidulans. Though I was not able to do so, I have discovered differences in telomeres between various strains of A. nidulans that rendered our anchored telomere PCR assay effective in one strain while ineffective in others in the process.
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