High-Throughput Genetic Analysis Using Microfabricated 96-Sample Capillary Array Electrophoresis Microplates

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 95; no. 5; pp. 2256 - 2261
• Main Authors: Simpson, Peter C., Roach, David, Woolley, Adam T., Thorsen, Todd, Johnston, Rick, Sensabaugh, George F., Mathies, Richard A.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 03.03.1998; National Acad Sciences; National Academy of Sciences; The National Academy of Sciences
• Subjects: Anodes; Biological Sciences; Capillaries; Capillary Action; Cathodes; Chemistry; DNA; Elastomers; Electrodes; Electrophoresis; Electrophoresis - instrumentation; Electrophoresis - methods; Equipment Design; Fluorescence; Gels; Genetics; Hemochromatosis - blood; Hemochromatosis - genetics; Hemochromatosis Protein; Histocompatibility Antigens Class I - biosynthesis; Histocompatibility Antigens Class I - genetics; HLA Antigens - biosynthesis; HLA Antigens - genetics; Humans; Membrane Proteins; Microchemistry - instrumentation; Microchemistry - methods; Physics; Point Mutation; Polymerase chain reaction; Polymerase Chain Reaction - instrumentation; Polymerase Chain Reaction - methods
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.95.5.2256

Capillary array electrophoresis (CAE) microplates that can analyze 96 samples in less than 8 min have been produced by bonding 10-cm-diameter micromachined glass wafers to form a glass sandwich structure. The microplate has 96 sample wells and 48 separation channels with an injection unit that permits the serial analysis of two different samples on each capillary. An elastomer sheet with an 8 by 12 array of holes is placed on top of the glass sandwich structure to define the sample wells. Samples are addressed with an electrode array that makes up the third layer of the assembly. Detection of all lanes with high temporal resolution was achieved by using a laser-excited confocal fluorescence scanner. To demonstrate the functionality of these microplates, electrophoretic separation and fluorescence detection of a restriction fragment marker for the diagnosis of hereditary hemochromatosis were performed. CAE microplates will facilitate all types of high-throughput genetic analysis because their high assay speed provides a throughput that is 50 to 100 times greater than that of conventional slab gels.