Identification of Point Mutations by Polymerase Chain Reaction/Ligase Detection Reaction/Flow-through Microarray with a Magnetic Bead-packed Capillary

Abstract

The polymerase chain reaction/ligase detection reaction (PCR/LDR) method is employed to identify the allelic composition of point mutations in genes. This involves amplifying copies of a target gene by primary PCR, followed by allele-specific LDR. However, polyacrylamide gel electrophoresis and DNA microarrays, which are complex and time-consuming methods, have commonly been used for the selective detection of LDR products. In this study, a low-density flow-through microarray was fabricated within one hour for the rapid and facile detection of LDR products. This microarray consisted of clusters of magnetic beads with probe DNA for LDR products immobilized on their surfaces and placed at constant intervals in a capillary tube. The LDR product solution was flowed through the flow-through microarray for only one minute for probe-target hybridization, enabling the detection of the targeted LDR products.