Date of Award

5-2010

Document Type

Thesis

Degree Name

Master of Science (MS)

Legacy Department

Chemistry

Advisor

Harcum, Sarah W

Committee Member

Christensen , Kenneth

Committee Member

Chumanov , George

Abstract

High quality, intact RNA is required for DNA microarray studies, cloning, and reverse transcriptase polymerase chain reaction (rt-PCR) analysis. There are several analytical methods used to assess the RNA quality. The RNA Integrity Number (RIN) from the Agilent Bioanalyzer is one quality control assay used to evaluate RNA. For recombinant Escherichia coli cultured under stressful conditions the RNA profiles obtained using the Agilent Bioanalyzer indicate RNA degradation; however, RNA obtained and purified in parallel from unstressed recombinant cultures indicate acceptable RNA values without significant degradation. We proposed that for stressed E. coli the RIN value is not necessarily indicative of RNA purification-related degradation but could be used as a tool to characterize and detect stressful culture conditions that target ribosomal RNA. Also, these results suggest the need for caution when assessing RNA quality based on ribosomal RNA abundance in stressed cells.

Share

COinS