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Separation method for nucleic acid and application thereof

A technology for nucleic acid separation and electrophoresis, applied in the field of bioengineering, to achieve low-cost separation, improved sensitivity and stability, high accuracy and stability

Inactive Publication Date: 2012-12-26
杜权 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Despite this possibility, the quantitative detection of low-concentration nucleic acids is still a huge technical challenge, and there is no mature and universal solution, especially for samples with no obvious sequence characteristics such as DNA residues in vaccines detection

Method used

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  • Separation method for nucleic acid and application thereof
  • Separation method for nucleic acid and application thereof
  • Separation method for nucleic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] Embodiment 1. Electrophoresis tank structure

[0126] 1. A separated two-chamber electrophoresis tank

[0127] see figure 1 , the electrophoresis tank includes a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5 and an agarose gel partition 7; Zone 4 and intermediate zone 5, anode 2 and cathode 3 are respectively located in these two zones.

[0128] 2. A separated two-chamber electrophoresis tank with selective nucleic acid separation components

[0129] see figure 2 , the electrophoresis tank includes a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5, an agarose gel partition 7 and a selective nucleic acid separation part 10; The two regions separated by the glue partition 7 are the anode region 4 and the middle region 5 respectively, and the anode 2 and the cathode 3 are respectively located in these two regions; the selective nucleic acid separation component 10 is located between the anode region and the...

Embodiment 2

[0142] Example 2. Nucleic acid concentration

[0143] use figure 1 The shown two-chamber electrophoresis cell separated by agarose gel concentrates the nucleic acid components in solution. The volume of the anode area of ​​the electrophoresis tank is 20mL, the volume of the middle area is 200mL, the width of the agarose strip is 1cm, and the top surface exceeds the liquid level height of the electrophoresis buffer by 0.5cm, forming a separation between the anode area and the middle area of ​​the electrophoresis tank .

[0144] 1. Take 20ug of purified pGL3 (Promega) plasmid, dissolve it in 200mL 0.5×TBE, and the final concentration of the plasmid is 100ng / mL;

[0145] 2. Slowly add 200mL of plasmid solution to the middle area of ​​the electrophoresis tank, and add 20mL of 0.5×TBE to the anode area of ​​the electrophoresis tank;

[0146] 3. If figure 1 Connect the power electrodes as shown, and stop the electrophoresis after 20 minutes of electrophoresis at a constant volta...

Embodiment 3

[0150] Example 3. Nucleic acid enrichment based on selective nucleic acid separation components

[0151] use figure 2 A two-chamber electrophoresis cell with selective nucleic acid separation components separated by agarose gel is shown to concentrate nucleic acid components in solution. The volume of the anode area of ​​the electrophoresis tank is 20mL, the volume of the middle area is 200mL, the width of the agarose strip is 1cm, and the top surface exceeds the liquid level height of the electrophoresis buffer by 0.5cm, forming a separation between the anode area and the middle area of ​​the electrophoresis tank . The selective nucleic acid separation component is an activated DEAE cellulose membrane, located in the nucleic acid transfer channel between the anode area and the middle area.

[0152] 1. Activation of DEAE cellulose membrane: Cut a piece of DEAE cellulose membrane (Schleicher & Schuell, NA-45) with a cross-sectional size suitable for the electrophoresis area,...

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Abstract

The invention provides a separation method for nucleic acid, concretely provides a method for selective separation of nucleic acid molecules in an electric field according to the charge properties of the nucleic acid molecules. The invention further provides a separation apparatus used in the method, and applications of the method and the apparatus in the separation of nucleic acid.

Description

technical field [0001] The present invention relates to the technical field of bioengineering, in particular to a nucleic acid separation method, device and application thereof. Furthermore, the present invention relates to a method, a device and a method for removing residual host cell nucleic acid and detecting residual host nucleic acid content in vaccine products. application. Background technique [0002] Vaccination is traditionally one of the important measures for the comprehensive prevention of infectious diseases, and the immune effect and safety of vaccines have attracted much attention. At present, many vaccines including recombinant (CHO cell) hepatitis B vaccine and Vero cell rabies vaccine are cell culture vaccines, and the pollution of host cell nucleic acid is inevitably mixed in the vaccine production process. If a protein vaccine mixed with host cell nucleic acid is used to immunize the human body, unpredictable consequences may occur, which may cause ins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00C07H21/00C07K1/26
Inventor 杜权杜军
Owner 杜权
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