By Kayla Mann
Faculty mentor: Dr. Ginny Morriss
Myotonic dystrophy (DM1) is a disease that causes muscle wasting and weakening. DM1 is caused by the dystrophia myotonia protein kinase gene that is encoded with the CTG repeat. DM1 has yet to be fully understood, and currently is incurable. This experiment is being explored to look at the differential expression of myokine genes in a mouse model of DM1 using qRT-PCR. RNA samples from mice that have been treated with 960 CUG expansion repeats will be used. Mice RNA samples were picked blindly and unbiased. These mice RNA samples were used to prepare cDNA using the applied biosystems cDNA synthesis kit and the DNase I Treatment Kit. A thermal cycler was used to process both kits. A master mix using SYBR green DNA binding dye will be used to then be able to run the qPCR and obtain results. The internal control that will be used to compare to the experimental group will be expressed as ActB. The experimental group will target the myokines; Ccl5, IL6, and Tnfsf10. The expected results will show the expression levels of the test subjects vs. the control subjects compared to one another. The qPCR is expected to show a good amount of these specific myokine genes in the RNA samples that are being used, compared to no expression in the internal control samples. Overall, the qPCR is supposed to show how much expression of the genes are showing up in each sample provided.
