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2004
LSU-HHMI Summer Undergraduate Research Program |
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 Laura
Mirch (Carroll College), Steven Soper, Chemistry
Detection of K-ras Point Mutations With High Clinical Value
for Colorectal Cancers Using Endonuclease V/AK16D Ligase and
Ligase Detection Reaction Assays
Our research investigates the benefits of utilizing PCR/Ligase
Detection Reactions (LDR) coupled with endonuclease V/AK16D
ligase treatment for the detection of known and unknown point
mutations within genomic DNA. The research focused specifically
on the K -ras gene, implicated in the tumorogenesis of certain
cancers including colorectal cancer. Identification of point
mutations within this gene could lead to early detection and
optimization of treatment options, as well as identification
those with a predisposition to develop colorectal cancer. An
LDR technique was utilized to identify specific, known mutations
within the gene sequence, while the endonuclease/ligase treatment
identified unknown mutations within this gene sequence. Capillary
electrophoresis and slab gel electrophoresis were employed to
distinguish the products of both EndoV/ligase and LDR based
on the fragments’ size differences. These methods have
been found to be highly sensitive and conducive to high throughput
applications. Further research should explore the potential
to transfer the gel electrophoresis separation to a microelectrophoretic
device with the potential for clinical applications.
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