An integrated slidable direct polymerase chain reaction-capillary electrophoresis microdevice for rapid Y chromosome short tandem repeat analysis

Abstract

We have developed an integrated slidable direct polymerase chain reaction-capillary electrophoresis (Direct PCR-CE) microdevice to analyze mini-Y chromosome short tandem repeat (mini-Y STR) typing without a DNA purification step. The Direct PCR has been recently commercialized as a simple DNA amplification technique, which does not require any sample preparation steps such as cell lysis, DNA extraction and purification for amplifying specific target genes. By implementing the advantages of Direct PCR on a microdevice, we can amplify target min-Y STR loci directly from human whole blood in a micro-reactor (2 mu L) and the amplicons were successively separated by micro-capillary electrophoresis. The utilization of a slidable plate enables us to manually control the fluid without use of microvalves and microtubes, and the functional units of sample injection, Direct PCR, and CE analysis were sequentially and independently performed by switching the slidable plate to each unit. On the integrated slidable Direct PCR-CE microdevice, we could complete monoplex and multiplex mini-Y STR typing directly using human whole blood within 1 h