Method for treating biomolecules and method for analyzing biomolecules

一种生物分子、加热处理的技术，应用在生物分子的处理以及分析领域，能够解决纳米孔阻塞等问题

Active Publication Date: 2019-06-04
HITACHI HIGH-TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The double helix structure of DNA and the high-order structure of single-stranded DNA are used to obtain a three-dimensional structure larger than the nanopore, which becomes a major obstacle when DNA passes through the nanopore, and sometimes the nanopore is blocked by these structures

Method used

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  • Method for treating biomolecules and method for analyzing biomolecules
  • Method for treating biomolecules and method for analyzing biomolecules
  • Method for treating biomolecules and method for analyzing biomolecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] A sample solution 1 was prepared by subjecting an aqueous solution having the following composition 1 to heat treatment and cooling treatment. Heat treatment and cooling treatment After maintaining at 95° C. for 15 minutes, it was rapidly cooled to 4° C. (cooling rate: 10° C. / second) and maintained at 4° C. for 5 minutes.

[0095] Composition 1: 4.0M tetramethylammonium salt, 0.1M Tris

[0096] configured with figure 1 In the sample introduction region 104 of the nanopore analyzer having the structure shown, the diameter of the nanopore was 1.1 to 1.8 nm when the closed current generated when passing through the nanopore 102 was measured. In addition, the blocking current was detected using a patch clamp amplifier (Axopatch 200B amplifiers, manufactured by Molecular Devices, Inc.). The blocking current was detected under the conditions of a sampling rate of 50kHz and an applied voltage of +300mV. From the obtained detection data, evaluations were carried out for "blo...

Embodiment 2

[0098] A sample solution 2 was prepared by performing the heat treatment and cooling treatment described in Example 1 on an aqueous solution having the following composition 2. In addition, except that the sample solution 2 was used instead of the sample solution 1, the sealing current was measured and evaluated in the same manner as in Example 1.

[0099] Composition 2: 4.0M tetraethylammonium chloride, 0.1M Tris

Embodiment 3

[0122] A sample solution 9 was prepared by performing the heat treatment and cooling treatment described in Example 1 on an aqueous solution having the following composition 5. In addition, except that the sample solution 9 was used instead of the sample solution 1, the sealing current was measured and evaluated in the same manner as in Example 1.

[0123] Composition 5: 1.7M Potassium Chloride, 1.5M Tetraethylammonium Chloride, 0.1M Tris

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Abstract

The purpose of the present invention is to provide a method for treating biomolecules and a method for analyzing biomolecules with which it is possible to effectively suppress the occlusion of nanopores. The present invention is a method for treating biomolecules for analysis in which nanopores are used, wherein the method includes a step for preparing a sample solution that includes ammonium cations represented by a prescribed formula and biomolecules in which at least a portion of the higher-order structure has been fused.

Description

technical field [0001] The invention relates to a processing method and an analysis method of biomolecules. Background technique [0002] The analysis technology of the base sequence of nucleic acid, which is a kind of biomolecule, can be applied to the detection of genetic diseases that cause genetic diseases, the evaluation of the effectiveness and side effects of pharmaceuticals, the detection of genetic mutations related to cancer diseases, etc. become very important. Nucleic acid base alignment can be, for example, a fluorescence detection device using capillary electrophoresis (Thermo Fisher Scientific, 3500 Genetic Analyzer), a fluorescence detection device for nucleic acid immobilized on a plate (Illumina Analysis was not performed by Biotech, HiSeq2500, etc. However, since these devices require expensive fluorescence detectors and fluorescent reagents, the cost of the experiment tends to increase. [0003] Therefore, a method of analyzing the base sequence by det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2527/107C12Q2527/119C12Q2527/125C12Q2565/607C12Q2565/631C12Q1/68G01N27/4473G01N27/44791G01N33/48721C12Q1/6839C12Q1/6806C12Q1/6832
Inventor 藤冈满后藤佑介
Owner HITACHI HIGH-TECH CORP
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