Integration of CE-SSCP in genetic analysis: fabrication and optimization of the polymer matrix

Genetic analysis has become an essential aspect in the field of biological research. Genetic material contains an abundance of information that can be harnessed, from the pathological study of diseases to the identification of algal species used for alternative fuel. The detection of genetic species and/or their mutated counterparts is an approach that still warrants improvement, however. Capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) offers a rapid and highly-reproducible means to detect alterations in genetic material. In this project, its implementation in the field of genetic studies as a way to detect mutations in hereditary diseases and identifying species via genetic coding was done by optimizing the molecular weight of the polymer medium. The polymer, polydimethyl acrylamide (PDMA), was fabricated and characterized at different polymer molecular weights. The fabricated polymer was then used in order to detect mutations in a model DNA and to genetically identify between two algae species. The polymer molecular weights of 3.4, 1.8, and 0.26 MDa were used in order to determine how varying polymer molecular weights affect the resolution of the data. The data shows that decreasing polymer molecular weight decreases the resolution of the peaks that resulted from the CE. It was found that all the mutations were consistently detected at 1.8 MDa polymer molecular weights, and genetic identification can be done with all three polymer weights. Thus for detection of mutations in a clinical setting, a recommended polymer weight of 1.8 MDa should be used. For identification between species of algae, lower polymer molecular weights can be used since it results in a shorter elution time.