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    Detection of K-ras Colorectal Point Mutations Using Endonuclease V/AK16DLigase and Ligase Detection Reaction Assays

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    2005_MirchL_THS_000755.pdf (7.336Mb)
    Author
    Mirch, Laura
    Advisor
    Marilyn Schendel; John Addis; Robert Swartout
    Date of Issue
    2005-04-01
    Metadata
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    URI
    https://scholars.carroll.edu/handle/20.500.12647/2941
    Title
    Detection of K-ras Colorectal Point Mutations Using Endonuclease V/AK16DLigase and Ligase Detection Reaction Assays
    Type
    thesis
    Abstract
    This research investigates the benefits of utilizing PCR/Ligase Detection Reaction multiplexing coupled with endonuclease V/AK16D ligase treatment in the detection of known and unknown point mutations within DNA. The research focused specifically on the K-ras gene, implicated in the development of certain cancers including colorectal cancer. Identification of point mutations within this gene could lead to early detection of cancer, as well as identify those with a predisposition to develop cancer. An LDR technique was utilized to identify specific, known mutations within the gene sequence, while the endonuclease/ligase treatment identified unknown mutations within the gene sequence. Capillary electrophoresis and slab gel electrophoresis were employed to distinguish the products of both EndoV/ligase and LDR, respectively, based on the fragments’ size differences. These methods have been found to be highly sensitive and have high throughput. Further research should explore the potential to transfer the gel electrophoresis separation to a microelectrophoretic device with the potential for clinical applications.
    Degree Awarded
    Bachelor's
    Semester
    Spring
    Department
    Life & Environmental Sciences
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    • Life and Environmental Sciences Undergraduate Theses

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