Publication Date

1-1-1998

Document Type

Dissertation/Thesis

First Advisor

Johnson-Wint, Barbara

Degree Name

B.S. (Bachelor of Science)

Legacy Department

Department of Biological Sciences

Abstract

Until recently, the measurement of mRNA levels was very time consuming and required specific parameters. With the advent of RT-PCR, mRNA fragments could be amplified and easily detected and measured. The amount of mRNA loaded into the reaction is proportional to the amount of product DNA. Here, we propose a new method for mRNA detection and quantification. The product of the amplification reaction is resolved electrophoretically on a polyacrylamide gel and then transferred to a positively charged nylon membrane. This membrane is then stained for DNA with a non-mutagenic dye. The membrane is converted to a visual image via a scanner. This image is then analyzed with the NIH Image program. The results show that this method is indeed of adequate sensitivity for comparisons of mRNA levels. Using this protocol, a linear relationship was shown between the amount of starting RNA and the amount of product DNA. This method is safer, less time consuming and at least as efficient as previously described methods.

Comments

Includes bibliographical references.

Extent

9 unnumbered pages

Language

eng

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

Media Type

Text

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