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Multicapillary electrophoresis device, and sample analysis method

An electrophoresis device and polycapillary technology, which are applied in the direction of material analysis, measurement device, and analysis material by electromagnetic means, and can solve problems such as difficult quantitative comparison of fluorescence intensity.

Pending Publication Date: 2022-01-14
HITACHI HIGH-TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, according to the methods of Patent Documents 1 and 2, it is also difficult to perform an accurate quantitative comparison of fluorescence intensities among a plurality of capillaries.

Method used

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  • Multicapillary electrophoresis device, and sample analysis method
  • Multicapillary electrophoresis device, and sample analysis method
  • Multicapillary electrophoresis device, and sample analysis method

Examples

Experimental program
Comparison scheme
Effect test

no. 1 approach

[0034] First, refer to figure 1 The schematic diagram of FIG. 1 will describe the structure of the polycapillary electrophoresis apparatus of the first embodiment. This polycapillary electrophoresis apparatus 1 has an apparatus main body 101 and a control computer 102 .

[0035] The device main body 101 is connected to a control computer 102 through a communication cable. An operator operates the control computer 102 to control each part of the device main body 101 , and the control computer 102 receives data detected by the photodetector 104 . The control computer 102 has a display as a data display screen for displaying received data. In addition, the control computer 102 may be incorporated in the apparatus main body 101 .

[0036] The device main body 101 further includes an arithmetic control circuit 103 , a photodetector 104 , a constant temperature bath 105 , a capillary array 106 , a light source 107 , and a light irradiation unit 108 .

[0037] The arithmetic contr...

Embodiment

[0077] The effect of the embodiment of the present invention was actually confirmed using the samples shown below.

[0078] (sample)

[0079] PowerPlex (registered trademark) 4C Matrix Standard (manufactured by Promega) was used as a standard product during wavelength calibration. As an actual sample, a product amplified by a PowerPlex (trademark) 16HS System (manufactured by Promega) using human genomic DNA provided by Promega as a template was used. Samples were prepared according to standard protocols recommended by Promega. In addition, in this experiment, both the standard substance and the actual sample were labeled with four kinds of phosphors (5-FAM, JOE, TMR, and CXR).

[0080] (Analysis process)

[0081] In capillary electrophoresis, in general, different actual samples are often run in each capillary. However, in this experiment, in order to clarify the effect of the present invention, the same actual sample was analyzed equally for all the capillaries. More sp...

Deformed example 1

[0087] Next, Modification 1 of the first embodiment will be described. In the first embodiment, the correction coefficient k(nX) was calculated using the data at the time of wavelength calibration (step S300), but in this modification 1, the fluorescent substance X is labeled for an arbitrary sample with a known concentration and subjected to electrophoresis. , using the data of the resulting fluorescence intensity to calculate the correction coefficient k(nX).

[0088] Let c(nX) be the concentration of DNA in a sample whose concentration is known for calculating the correction coefficient k(nX). Here, n is the end number of the capillaries 119-1 to 4, and X is the type of phosphor. In addition, the average value obtained by averaging the DNA concentrations c(nX) among a plurality of capillaries was defined as avg(X). In addition, the concentration ratio r(nX) of DNA among a plurality of capillaries is defined as r(nX)=avg(X) / c(nX).

[0089] In each of the plurality of capi...

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Abstract

A multicapillary electrophoresis device is provided with a capillary array in which a plurality of capillaries are arranged, a light source that irradiates the plurality of capillaries with excitation light, a light detector that detects fluorescence from a sample within the capillaries, and a computation control unit that calculates the signal intensity of the fluorescence in accordance with a signal from the light detector. The computation control unit is configured so as to correct the signal intensity in accordance with a correction index derived for each combination of any of the capillaries and a fluorescent body marking the sample.

Description

technical field [0001] The invention relates to a multi-capillary electrophoresis device and a sample analysis method. Background technique [0002] Electrophoresis is widely known as a method for analyzing the nucleotide sequence or nucleotide length of DNA. In addition, as one type of electrophoresis method, there is a capillary electrophoresis method in which electrophoresis is performed in a capillary. In this capillary electrophoresis method, a sample containing DNA is injected into a capillary filled with a separation medium, and a high voltage is applied to both ends of the capillary in this state. At this time, DNA, which is a negatively charged particle, moves to the anode side in the capillary depending on its own size, and as a result, bands according to molecular weights are generated in the capillary. Each DNA is fluorescently labeled and irradiated with excitation light to emit fluorescence. Multiple fluorochromes are also sometimes used. By detecting this ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/447G01N27/44721G01N21/6428G01N21/6486G01N27/44782
Inventor 隅田周志穴泽隆藤冈满山崎基博中泽太朗
Owner HITACHI HIGH-TECH CORP