RESUMO
An integrated microfluidic device capable of performing a variety of genetic assays has been developed as a step towards building systems for widespread dissemination. The device integrates fluidic and thermal components such as heaters, temperature sensors, and addressable valves to control two nanoliter reactors in series followed by an electrophoretic separation. This combination of components is suitable for a variety of genetic analyses. As an example, we have successfully identified sequence-specific hemagglutinin A subtype for the A/LA/1/87 strain of influenza virus. The device uses a compact design and mass production technologies, making it an attractive platform for a variety of widely disseminated applications.
Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Influenza Humana/genética , Técnicas Analíticas Microfluídicas/instrumentação , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Animais , Primers do DNA/química , DNA Viral/metabolismo , Eletroforese , Vidro , Temperatura Alta , Humanos , Processamento de Imagem Assistida por Computador , Camundongos , Microfluídica , Miniaturização , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Silício/química , Temperatura , Fatores de TempoRESUMO
Microfabrication techniques have become increasingly popular in the development of the next generation of DNA analysis systems. While significant progress has been reported by many researchers, complete microfabricated integrated DNA analysis devices are still in the earliest stages of development. Most miniaturized analysis systems have incorporated noncross-linked polymer solutions as the separation medium of choice and the operation of these systems necessitates the use of high electric fields and long separation lengths. In this paper, we present two techniques that may help alleviate this problem and accelerate the development of the so-called 'lab-on-a-chip' systems. We present the use of photodefinable polyacrylamide gels as a sieving medium for DNA electrophoresis. These gels offer the significant advantages of faster curing times, locally controlled gel interface, and simpler handling over chemically polymerized gels. We also introduce an electrode-defined sample compaction and injection technique. This technique helps achieve sample compaction without migration into the gel and offers significant control over the size and application of the sample plug. The use of these technologies for double-stranded DNA separations in microfabricated separation systems is demonstrated.
