Amplification and separation of nucleic acid sequences using strand displacement amplification and bioelectronic microchip technology
First Claim
1. A method for the amplification of one or more target nucleic acids of interest in one or more samples using a bioelectronic microchip, comprising:
- a) introducing at least one of the target nucleic acids onto a bioelectronic microchip having a plurality of electronically addressable capture sites;
b) electronically addressing the target nucleic acid to at least one capture site which has attached thereto at least a first PCR oligonucleotide primer, wherein the first PCR primer comprises a sequence specific for the target nucleic acid to be amplified;
c) hybridizing the target nucleic acid to be amplified to the first PCR primer at the capture site;
d) contacting the hybridized target nucleic acid with enzymes and reagents necessary to support PCR, including a DNA polymerase activity; and
e) amplifying the target nucleic acid by PCR to produce amplicon species.
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Accused Products
Abstract
Described and disclosed are devices, methods, and compositions of matter for the multiplex amplification and analysis of nucleic acid sequences in a sample using novel strand displacement amplification technologies in combination with bioelectronic microchip technology. Specifically, a nucleic acid in a sample is amplified to form amplicons, the amplicons are addressed to specified electronically addressable capture sites of the bioelectronic microchip, the addressed amplicons are captured and labeled, and then the capture sites are analyzed for the presence of label. Samples may be amplified using strand displacement amplification. The invention is also amenable to other amplification methodologies well known by those skilled in the art. The capture and label steps may be by a method of universal capture with sequence specific reporter, or by a method of sequence specific capture with universal reporter. The label may be detected by fluorescence, chemiluminescence, elecrochemiluminescence, or any other technique as are well known by those skilled in the art. This invention further allows for analyzing multiple nucleic acid targets on a single diagnostic platform wherein the nucleic acids may be amplified while either in direct contact with microchip components or in solution above the microchip array.
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Citations
33 Claims
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1. A method for the amplification of one or more target nucleic acids of interest in one or more samples using a bioelectronic microchip, comprising:
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a) introducing at least one of the target nucleic acids onto a bioelectronic microchip having a plurality of electronically addressable capture sites;
b) electronically addressing the target nucleic acid to at least one capture site which has attached thereto at least a first PCR oligonucleotide primer, wherein the first PCR primer comprises a sequence specific for the target nucleic acid to be amplified;
c) hybridizing the target nucleic acid to be amplified to the first PCR primer at the capture site;
d) contacting the hybridized target nucleic acid with enzymes and reagents necessary to support PCR, including a DNA polymerase activity; and
e) amplifying the target nucleic acid by PCR to produce amplicon species. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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Specification