A kind of preparation method of low solidification temperature agarose

A low coagulation 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 coagulation temperature and Melting temperature, low cost effect

Active Publication Date: 2018-02-23
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 of agarose powder in 150mL of deionized water at 90°C to prepare an agarose solution;

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

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

[0034] (4) The solution of step (3) is rapidly cooled to 5° C., slowly add 20 to 60% (v / v) ethylene oxide aqueous solution dropwise in 60 min while stirring until the concentration of ethy...

Embodiment 2

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

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

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

[0043] (4) Rapidly cool the solution in step (3) to 5°C, and slowly add 20-60% (v / v) ethylene oxide aqueous solution dropwise within 60 minutes w...

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-mentioned agarose solution is naturally cooled to 82°C, 750mL of sodium borohydride sodium hydroxide solution is added to it for a reduction reaction for 15 minutes. During the reaction, the color change of the solution is observed. When the color changes from light green to a colorless transparent liquid , terminate the reduction reaction, in the sodium hydroxide solution of sodium borohydride, the concentration of sodium borohydride is 4.4mol / L, and the solvent is 10.0~20.0mol / L NaOH solution;

[0051] (3) Adding 30L concentration to the solution obtained in step (2) is that the sodium hydroxide solution of 3.8mol / L is carried out alkalization treatment, and replenishes 60L deionized water;

[0052] (4) Rapidly cool the solution in step (3) to 5°C, and slowly add 20-60% (v / v) ethylene oxide aqueous solution dropwise within 60 minutes while s...

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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 in particular relates to a preparation method of low solidification temperature agarose. Background technique [0002] Agarose is isolated from red algae agar, and is a polysaccharide copolymer that is alternately linked by β~D~galactopyranose and 3,6~dehydration~α~L~galactopyranose. Agarose contains less sulfate radicals than agar, has special gelling properties, especially remarkable stability and hysteresis, and is easy to absorb water and has a special stabilizing effect. It has been widely used in biology, food, medicine, chemical industry, textile , national defense and other fields. [0003] The coagulation temperature of the agarose gel obtained by direct separation of natural red algae agar is generally 35-40°C. At this temperature, many organisms or heat-sensitive reagents will be inactivated, so it is necessary to prepare agarose with a lower coagulation temperature. ...

Claims

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Application Information

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Patent Type & Authority Patents(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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