B.S. (Bachelor of Science)
Department of Biological Sciences
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.
Cancasci, Vince J., "A novel method for the detection and the quantification of RT-PCR products" (1998). Honors Capstones. 47.
9 unnumbered pages
Northern Illinois University
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