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Preparation method of low-solidification-temperature agarose

A low solidification temperature, agarose technology, applied in the field of agarose preparation, can solve the problems of agarose gel strength measurement, low product gel strength, modifier toxic substances, etc., to achieve good product quality, low solidification temperature and Melting temperature, low cost effect

Active Publication Date: 2016-05-04
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

People such as Guiseley use dimethyl sulfate, propylene oxide, halogenated alkanes to modify, but do not measure the gel strength of the modified agarose, only draw the relationship between modifier addition, degree of substitution, and solidification temperature. relation
[0005] To sum up, the main problem of the above-mentioned modification at present is that most of the modifiers used are toxic substances, there are safety hazards in the production, and there are also problems such as the gel strength of the product is too low and the solidification temperature is too high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Dissolve 4.0g agarose powder in 150mL deionized water at 90°C to prepare an agarose solution;

[0032] (2) After the above agarose solution is naturally cooled to 82°C, add 0.9 mL of sodium borohydride and sodium hydroxide solution to it for a reduction reaction for 15 minutes. During the reaction, observe the color change of the solution. When the color changes from light green to colorless and transparent When liquid, stop the reduction reaction. In the sodium hydroxide solution of sodium borohydride, the concentration of sodium borohydride is 4.4 mol / L, and the solvent is 10.0-20.0 mol / L NaOH solution;

[0033] (3) Add 40 mL of sodium hydroxide solution with a concentration of 3.8 mol / L to the solution obtained in step (2) for alkalization;

[0034] (4) Cool the solution of step (3) quickly to 5°C, and slowly add 20-60% (v / v) ethylene oxide aqueous solution dropwise within 60 minutes while stirring until the concentration of ethylene oxide in the solution is 1.0~5.0wt%...

Embodiment 2

[0040] (1) Dissolve 30g agarose powder in 575mL deionized water at 90°C to prepare an agarose solution;

[0041] (2) After the above agarose solution is naturally cooled to 82°C, add 6.8mL sodium borohydride and sodium hydroxide solution to it for reduction reaction for 15 minutes. During the reaction, observe the color change of the solution. When the color changes from light green to colorless and transparent When liquid, stop the reduction reaction. In the sodium hydroxide solution of sodium borohydride, the concentration of sodium borohydride is 4.4 mol / L, and the solvent is 10.0-20.0 mol / L NaOH solution;

[0042] (3) Add 300 mL of sodium hydroxide solution with a concentration of 4.0 mol / L to the solution obtained in step (2) for alkalization, and add 575 mL of deionized water;

[0043] (4) Cool the solution of step (3) quickly to 5℃, and slowly add 20-60% (v / v) ethylene oxide aqueous solution to the concentration of ethylene oxide in the solution within 60 minutes while stirrin...

Embodiment 3

[0049] (1) Dissolve 3kg of agarose powder in 60L of deionized water at 90°C to prepare an agarose solution;

[0050] (2) After the above agarose solution is naturally cooled to 82°C, add 750mL of sodium borohydride and sodium hydroxide solution to the reduction reaction for 15 minutes. During the reaction, observe the color change of the solution. When the color changes from light green to colorless transparent liquid When the reduction reaction is terminated, the concentration of sodium borohydride in the sodium hydroxide solution of sodium borohydride is 4.4 mol / L, and the solvent is 10.0-20.0 mol / L NaOH solution;

[0051] (3) Add 30L of sodium hydroxide solution with a concentration of 3.8mol / L to the solution obtained in step (2) for alkalization, and add 60L of deionized water;

[0052] (4) Cool the solution of step (3) quickly to 5°C, and slowly add 20-60% (v / v) ethylene oxide aqueous solution to the concentration of ethylene oxide in the solution within 60 minutes while stirri...

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Abstract

The invention discloses a preparation method of low-solidification-temperature agarose. The preparation method includes the following steps that 1, agarose is dissolved in deionized water to prepare an agarose solution; 2, the agarose solution is naturally cooled to 60-83 DEG C, and then a sodium hydroxide solution of sodium borohydride is added into the agarose solution for a reduction reaction; 3, a sodium hydroxide solution is added for alkalization treatment; 4, rapid cooling is carried out, and oxirane is slowly dripped with stirring for a derivatization reaction; 5, heating is carried out, and an acetic acid solution is dripped with stirring to regulate the pH value of the solution; 6, hot isopropanol is added with stirring for alcohol precipitation fractionation; 7, filtering, smashing and washing are carried out several times to obtain a filter cake; 8, after drying, smashing and screening, low-solidification-temperature agarose is obtained. According to the preparation method, common agarose is subjected to chemical modification and fractionation purification, and prepared agarose has the advantages of being low in solidification temperature and melting temperature, high in jelly strength and the like and is an ideal biological material for DNA separation and extraction.

Description

Technical field [0001] The invention belongs to the technical field of agarose preparation, and specifically relates to a method for preparing agarose with a low solidification temperature. Background technique [0002] Agarose is separated from red algae agar, which is a polysaccharide copolymer composed of β~D~galactopyranoside and 3,6~anhydro~α~L~galactopyranoside alternately connected. Agarose contains less sulfate radicals than agar, and has special gelling properties, especially significant stability and hysteresis, and it is easy to absorb water, and has a special stabilizing effect. It has been widely used in biology, food, medicine, chemicals, and textiles. , National defense and other fields. [0003] The solidification temperature of the agarose gel obtained by the direct separation of natural red algae agar is generally 35-40℃. At this temperature, many organisms or heat-sensitive reagents will be inactivated, so it is necessary to prepare agarose with a lower solidifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/12
CPCC08B37/12
Inventor 肖美添张学勤叶静黄雅燕赵鹏王江林
Owner HUAQIAO UNIVERSITY
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