Method and device for performing submicroliter reactions with nucleic acids or proteins

A nucleic acid, reaction technique used in the field of preparing and conducting small-scale reactions using nucleic acids or proteins

Inactive Publication Date: 2006-06-21
AMERSHAM BIOSCI SV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, such methods require specialized equipment and a good familiarity with mass spectrometers

Method used

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  • Method and device for performing submicroliter reactions with nucleic acids or proteins
  • Method and device for performing submicroliter reactions with nucleic acids or proteins
  • Method and device for performing submicroliter reactions with nucleic acids or proteins

Examples

Experimental program
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Effect test

Embodiment 1

[0262] Reaction Mixture Preparation Example 1: Reagents are pipetted and mixed on a substrate using a capillary cartridge

[0263] One way to prepare the reaction mixture is to use a pipette to measure the components of the reaction mixture separately. For example, when preparing a PCR mix, the nucleic acid sample and the PCR reagents will be measured separately and then dispensed into the same container where they are brought together. When using the automated system of FIG. 1 , an automated robot 102 moves a transfer head 104 containing capillary cassettes to the position where the sample plate is located. Dip the capillary end of the capillary cartridge into the well. The capillary is primed by capillary action to measure precise amounts of sample. The wells of the sample plate contain PCR-amplified nucleic acid samples. DNA samples should be diluted sufficiently so that each capillary in the capillary cartridge contains 5-20 ng of DNA in a volume of 10-10,000 nL.

[02...

Embodiment 2

[0273] Example 2 of Capillary Cartridge Sampling: Gas Replacement

[0274] A second method of dividing the liquid contained in the capillary tubes of the capillary cartridge is to use a gas displacement device. As shown in FIG. 1 , the microplates from the microplate residence chamber are transferred by the transfer head 104 to the sample dispensing device position 122 . A gas sample divider is installed here, such as the sample divider depicted in Figures 4A-C. The transfer head 104 then lifts one more capillary cartridge and primes it with the sample from the multiwell plate or with the reagents. The capillary cartridge moves to the sample dispensing device position 122 and contacts the gas displacing device head. The substrate of the capillary cartridge is placed on the receiving platform on the head of the gas displacement device. In addition, the gas displacement device head can be connected to the automated robot 102 .

[0275]As shown in FIG. 4A , a gas displacement...

Embodiment 3

[0284] Reaction Mixture Assembly Example 3: Solid Phase Capture

[0285] A third method of assembling the reaction mixture is to capture species from the sample on the substrate surface, such as the inner surface of a capillary segment. The captured species can be nucleic acids, enzymes, other biopolymers or chemicals. Desired species captured from a sample can be attached to surfaces in a variety of ways. These methods include covalent binding, antibody binding, DNA hybridization, hydrophobic interactions, electric fields, magnetic fields or other chemical or physical forces. After the sample is bound, the liquid remaining in the suspended sample can be drained from the capillary or microchip by chemical reaction or physical force. The capillary can be used for gas displacement or centrifugation, vacuum method can also be used. Sample material will remain on the surface of the substrate. In this single step, the sample material is also substantially purified. The reactio...

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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.

Description

[0001] This invention was made with government support. The government has certain rights in this invention. [0002] Cross-references to related applications [0003] This application claims U.S. Provisional Application No. 60 / 355,660, filed February 8, 2002, U.S. Provisional Application No. 60 / 355,648, filed February 8, 2002, and U.S. Provisional Application No. Patent application 10 / 262,476 enjoys priority. field of invention [0004] The invention belongs to the field of biotechnology, and relates to methods and devices for preparing and performing small-scale reactions using nucleic acids or proteins. Background of the invention [0005] The federally funded Human Genome Project initially aimed to sequence all ten folds of the human genome by 2005. With the pace greatly accelerated, there have been partial sketches recently. However, this feat did not reduce the demand for fast, cheap DNA sequencing, but instead stimulated the demand for fast, cheap nucleic acid seq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34
Inventor S·B·约万诺维奇O·萨拉斯-索拉诺李政勳
Owner AMERSHAM BIOSCI SV
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