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Preparation method of agarose with low electroendosmosis

A technique of agarose and electro-endosmosis, which is applied in the field of biochemistry, can solve problems such as inapplicability, and achieve the effects of low electro-endosmosis value, improved value and simple preparation method.

Inactive Publication Date: 2014-07-09
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Another literature report uses the DEAE-cellulose resin method to prepare the agarose from Geliflower agar with an electroosmotic value of 0.12, but this value is still relatively high; secondly, only using the DEAE-cellulose resin method is not suitable for other red algae source of agar

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Isolation and preparation of primary agarose from agar

[0034] Weigh 4g of agar powder (Agar agar) and add it to 200mL of pH4.4 disodium hydrogen phosphate-citric acid buffer.

[0035] (Disodium hydrogen phosphate-citric acid buffer solution, pH4.4 disodium hydrogen phosphate-citric acid buffer solution preparation method:

[0036] A----preparation of 0.2mol / L disodium hydrogen phosphate solution: weigh disodium hydrogen phosphate dodecahydrate (Na 2 HPO 4 .12H 2 O--molecular weight 358.14) 71.63g, dissolved in distilled water, and dilute to 1L.

[0037] B----Preparation of 0.1mol / L citric acid solution: Weigh 42.03g of citric acid monohydrate (molecular weight: 210.14), dissolve it in distilled water, and set the volume to 2L.

[0038] C----Measure 882ml of 0.2mol / L disodium hydrogen phosphate solution and 1118ml of 0.1mol / L citric acid solution respectively, and mix them to obtain pH 4.4 disodium hydrogen phosphate-citric acid buffer), keep the temperature at ...

Embodiment 2

[0043] 1. Isolation and preparation of primary agarose from agar

[0044] Weigh 12g of agar powder (gracilaria agar) and add it to 200mL of pH4.4 disodium hydrogen phosphate-citrate buffer solution, and stir at a constant temperature of 50°C at a speed of 20 rpm for 18 hours. Filtrate while hot and collect the filtrate. Wash with distilled water repeatedly until the filtrate is neutral, and then wash twice with ultrapure water. Put it in a 50°C electric constant temperature blast drying oven to dry, weigh, and pulverize to obtain 9.23 g of primary agarose, with a yield of 76.9%. It was determined that the electroosmosis value was 0.20.

[0045] 2. Separation and preparation of low-voltage endosmotic grade agarose from agar

[0046] Weigh 6.0g of primary agarose prepared by disodium hydrogen phosphate-citric acid buffer method, add 600ml of distilled water, heat to dissolve, add 5g of pretreated DEAE-cellulose resin (soaked in 10%NaCl for 20h, 10%HCl Soak for 4h, soak in 4%...

Embodiment 3

[0048] 1. Isolation and preparation of primary agarose from agar

[0049] Weigh 20g of agar powder (gracilaria agar) and add it to 200mL of pH4.4 disodium hydrogen phosphate-citrate buffer solution, and stir at a constant temperature of 40°C at a speed of 15 rpm for 24 hours. Filtrate while hot and collect the filtrate. Wash with distilled water repeatedly until the filtrate is neutral, and then wash twice with ultrapure water. Put it in a 55°C electric constant temperature blast drying oven to dry, weigh, and pulverize to obtain 15.45 g of primary agarose, with a yield of 77.25%. It was determined that the electroosmosis value was 0.21.

[0050]2. Separation and preparation of low-voltage endosmotic grade agarose from agar

[0051] Weigh 10g of primary agarose prepared by disodium hydrogen phosphate-citric acid buffer method, add 1000ml of distilled water, heat to dissolve, add 25g of pretreated DEAE-cellulose resin (soaked in 10%NaCl for 20h, 10%HCl in turn 4h, soaked in...

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Abstract

The invention discloses a preparation method of agarose with low electroendosmosis. The preparation method comprises the following steps of A, adding agar powder to disodium hydrogen phosphate-citric acid buffer solution with pH of 4.4, wherein the addition amount of the agar powder is 20-100g / L; B, stirring; C, carrying out hot suction filtration and collecting filtrate; D, repeatedly washing the filtrate with distilled water to be neutral; E. drying and crushing to obtain primary agarose; F, adding 1 part by weight of primary agarose to 150-250 parts by weight of distilled water, and heating for dissolving; G. adding 3-7 parts by weight of pretreated DEAE-cellulosic resin; H, carrying out hot suction filtration and collecting filter residue; I, washing the filter residue with high temperature distilled water of more than 85 DEG C, and collecting filtrate; J, drying the filtrate and crushing to obtain agarose with low electroendosmosis. The preparation method has the advantages that both the used raw materials and the preparation method are simple; the obtained agarose has low electroendosmosis value which can be below 0.1.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a method for preparing low-electricity endosmosis agarose. Background technique [0002] Electroendosmosis is one of the important quality standards of agarose, which to some extent reflects the negative charge of the agarose gel. Electroendosmosis has a great influence on electrophoresis, especially on isoelectric focusing electrophoresis. When agarose is used as the electrophoresis medium, negatively charged groups such as sulfate groups and acetonyl groups contained in agarose make the agarose gel negatively charged. During electrophoresis, although the gel itself does not move to the positive electrode under the action of the electric field force, the water in the gel moves to the cathode under the action of hydronium ions, and the neutral molecules in the sample also move to the cathode along with the water. Electro-endosmosis complicates the electrophoresis process, and more i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/12
Inventor 林毅班珍
Owner HUAQIAO UNIVERSITY
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