Methods and apparatus for performing submicroliter reactions with nucleic acids or proteins
First Claim
1. A method of performing an enzymatic reaction in a capillary tube using a normalized quantity of a nucleic acid, comprising:
- saturably binding a nucleic acid from an excess thereof directly on an inner surface of said capillary tube by contacting said inner surface with a solution comprising said nucleic acid and a chaotropic agent for a time sufficient for the nucleic acid to become saturably bound to said inner surface;
removing unbound excess of the nucleic acid therefore generating a normalized quantity of said nucleic acid;
introducing an enzymatic reaction mixture into said capillary tube having said normalized quantity of said nucleic acid;
wherein said reaction mixture comprises an oligonucleotide primer, a DNA polymerase, dNPTs, and at least one dideoxynucleotide triphosphate (ddNTP),performing said enzymatic reaction in said capillary tube using said normalized quantity of said nucleic acid.
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Abstract
Methods for preparing nanoscale reactions using nucleic acids or proteins are presented. Nucleic acids are captured saturably, yet reversibly, on the internal surface of the reaction chamber, typically a capillary. Excess nucleic acid is removed and the reaction is performed directly within the capillary. Proteins are captured specifically and saturably on the modified inner surface of the reaction chamber, typically a capillary. Excess protein is removed and the reaction is performed directly within the capillary. Devices for effecting the methods of the invention and a system designed advantageously to utilize the methods for high throughput reactions involving nucleic acids or proteins are also provided.
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Citations
35 Claims
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1. A method of performing an enzymatic reaction in a capillary tube using a normalized quantity of a nucleic acid, comprising:
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saturably binding a nucleic acid from an excess thereof directly on an inner surface of said capillary tube by contacting said inner surface with a solution comprising said nucleic acid and a chaotropic agent for a time sufficient for the nucleic acid to become saturably bound to said inner surface; removing unbound excess of the nucleic acid therefore generating a normalized quantity of said nucleic acid; introducing an enzymatic reaction mixture into said capillary tube having said normalized quantity of said nucleic acid; wherein said reaction mixture comprises an oligonucleotide primer, a DNA polymerase, dNPTs, and at least one dideoxynucleotide triphosphate (ddNTP), performing said enzymatic reaction in said capillary tube using said normalized quantity of said nucleic acid. - 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, 34, 35)
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Specification