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Analysis of Nucleosome Isolation and Recovery: From <em>In Silico</em> Invitrosomes to <em>In Vivo</em> NucleosomesSkousen, Collin Brendan 01 December 2016 (has links)
There are a vast number of factors that influence nucleosome formation, and consequently gene regulation. These factors include histone modifications, nucleotide composition, transcriptional region elements, and specific nucleotide motifs, among others. Although the amount we know now is limited, we are creating new techniques and discoveries to assist us in continued understanding of chromatin. To make a significant contribution to the field of chromatin, I conducted two hypothesis driven sets of experiments that address the topic of chromatin structure. First, I created a technique for tissue specific nucleosome isolation with the goal of observing the effect of single nucleotide polymorphisms (SNPs) on nucleosome formation. Second, I created and tested a method to recover lost in vitro nucleosome reconstitution data, which can improve this type of data, commonly used for observing nucleosome positioning. The first experiment needs a more specific antibody to complete the last step and function as designed. The second experiment shows that our nucleosome recovery method, when applied conservatively, can recover 90% of the lost nucleosome data.
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