Electrophoretic buffer

A buffer and electrophoresis technology, which is applied in material analysis, nanotechnology, immunoassay, etc. by electromagnetic means, can solve problems such as difficult injection, time-consuming, and sensitivity of buffer viscosity to temperature.

Inactive Publication Date: 2005-07-20
JAPAN SCI & TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Existing buffers for electrophoresis have excellent performance in terms of separation ability for limited sizes, but due to the use of high-viscosity polymers (methylcellulose, hydroxycellulose, polyacrylamide, etc.), there are 1) It takes time to inject the buffer into the microchannel, 2) as the channel width narrows, the injection becomes difficult, 3) it is necessary to change the concentration of the polymer according to the size of the DNA, and when separating a wide range of DNA sizes, the resolution There are limitations, 4) the viscosity of the buffer is sensitive to temperature and other shortcomings

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0133] modulation polymeric micelles

[0134] Make ethylene glycol 130mmol, 6.5ml and dry THF 30ml, 2-methoxyethanol 0.16ml, naphthalene potassium (Naftarencarium) 1mmol polymerize at 25°C for two days, and then add 3,6-dimethyl 1,4 - 32 mmol and 31 ml of dioxane-2,5-dione were polymerized together at 25° C. for 2 hours, and then 20 mmol and 3.6 mL of methacrylic anhydride were used to terminate the reaction. Purification by reprecipitation was carried out in 600 mL of cold isopropanol, precipitation was carried out by centrifugation, and the product was recovered. This product was dissolved in 100 mL of benzene, freeze-dried and recovered to obtain 2.5 g of a copolymer. 2.5 g of the obtained copolymer was dissolved in 1000 mL of water, kept at 80° C. in an oil bath, heated while stirring for 6 hours, and left overnight after heating to recover the micellar solution. After 30 minutes, thermal polymerization was performed for 20 hours while maintaining 10,000 mL of the obtain...

example 1

[0154] Example 1 (polymer micelles + common method)

[0155] Using high molecular weight micelles as the buffer for electrophoresis, electrophoresis was performed according to the ordinary method, and the two DNA markers of 100 bp and 800 bp were separated. As a result, no peak appeared within the migration time of 180s. ( figure 1 )

example 2

[0156] Example 2 (polymer micelles + PP method)

[0157] According to the PP method, the operation in Example 1 was carried out. Let the P intensity of the former be M, and the P intensity of the latter be L. As a result, two peaks were seen within the migration time of 180s. ( figure 2 )

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Abstract

By using an electrophoretic buffer comprising a polymerized polymer micelle formed by the steps comprising dispersing into an aqueous medium a block copolymer represented by HPLS-HPBS-PLZA, wherein HPLS is a hydrophilic polymer segment, HPBS is a hydrophobic polymer segment, and PLZA is a polymerizable group having an ethylenically unsaturated double bond, and polymerizing the block copolymer as a buffer for a capillary electrophoresis or a microchip electrophoresis, pressurizing the sample after introduction at a given pressure for a given time period, and electrophoresing in an electrophoretic electric field, a polymer compound such as DNA can be separated rapidly and in high separation ability.

Description

technical field [0001] The present invention relates to a buffer for electrophoresis and an electrophoresis method capable of rapidly separating high-molecular compounds with high separation ability. Background technique [0002] Existing buffers for electrophoresis have excellent performance in terms of separation ability for limited sizes, but due to the use of high-viscosity polymers (methylcellulose, hydroxycellulose, polyacrylamide, etc.), there are 1) It takes time to inject the buffer into the microchannel, 2) as the channel width narrows, the injection becomes difficult, 3) it is necessary to change the concentration of the polymer according to the size of the DNA, and when separating a wide range of DNA sizes, the resolution There are limitations, 4) the viscosity of the buffer solution is sensitive to temperature and other shortcomings. Therefore, a buffer solution for electrophoresis that does not have the above-mentioned problems 1) to 4) and can separate polyme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCY10T436/108331G01N27/44747B82B1/00G01N27/447
Inventor 马场嘉信片冈一则田渕真理长崎幸夫田中靖子桑原智枝
Owner JAPAN SCI & TECH CORP
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