A Novel Approach Using the UBe3A and TSC2 Genes
Genotyping is a common process used to determine differences between the genetic makeup of DNA sequences. Through the use of biological assays and other methods, the length of DNA strands can be visualized which will differentiate the condition of the subject. Currently, genotyping can be a faulty process that leads to inaccurate results due to contaminants and denigration of DNA.
After my first experience genotyping, I wanted to find a solution to this by finding a way to produce clearer images in genotyping, despite possible contaminants. In this project, the UBE3A and TSC2 genes were tested using wild type and mutated type samples to show different chromosomal patterns.
By adding variances in time, temperature, and Proteinase K in genotyping solutions, the effects of each variable were identified and the best combination was found to solve the problem of unclear genotyping images.
Mixtures with Proteinase K proved to be significantly clearer than those without; room temperature provided more intense pixel saturation for TSC2 and cold temperature provided more intense pixel saturation for UBE3a. Clarity seemed to degrade over time.
The results conclude that scientists should add Proteinase K in their genotyping mixtures for clearer images. In both TSC2 and UBE3A mice, the effects of Time and Condition were significant; however, only in TSC2 mice was condition by time significant.
These results can apply to labs around the world and will significantly improve the accuracy in scientists’ genotyping data.