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Pellosil activate fluid and applications thereof for extracting nucleic acid

A technology of silica gel membrane and activation solution, which is applied in organic chemistry, chemical instruments and methods, DNA preparation, etc., can solve the problems of silica gel membrane instability, decline, and inhomogeneous nucleic acid binding ability, and achieve good uniformity and stability. The effect of enhanced binding capacity

Inactive Publication Date: 2009-01-07
TIANGEN BIOTECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] Direct use of silica gel membrane for nucleic acid separation has disadvantages such as instability, inhomogeneity, and decreased binding ability to nucleic acid. Our special activation process can overcome the above defects

Method used

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  • Pellosil activate fluid and applications thereof for extracting nucleic acid
  • Pellosil activate fluid and applications thereof for extracting nucleic acid
  • Pellosil activate fluid and applications thereof for extracting nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Silica gel membrane activation process

[0052] Cut the silica gel membrane with a thickness of about 0.3 mm for air filtration into an appropriate size and place it in the plastic adsorption column CB3 together with the filter gasket, add 500 μl of silica gel membrane to the adsorption column CB3 (the adsorption column is placed in a 2ml collection tube) to activate centrifuge at 12,000rpm for 30-60 seconds, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube. (Columns treated with activation solution are best used immediately).

[0053] The composition of the activation solution is: 100mM NaHCO 3 , 5% (w / v) SDS, 25 mM EDTA.

[0054] 2. Extract plasmid pcDNA with activated silica membrane

[0055]Take 1-3ml of overnight cultured E.coli (containing plasmid pcDNA) bacterial solution into a centrifuge tube, centrifuge at room temperature at 12,000rpm for 1 minute to collect the bacteria, and aspirate the super...

Embodiment 2

[0063] The silica gel membrane activation process is the same as in Example 1, except that the activation solution consists of: 50mM NaHCO 3 , 1% (w / v) SDS, 1 mM EDTA.

Embodiment 3

[0065] The silica gel membrane activation process is the same as in Example 1, except that the activation solution consists of: 200mM NaHCO 3 , 10% (w / v) SDS, 50 mM EDTA.

[0066] Experiments have proved that the binding ability of the silica gel membrane to nucleic acid after being treated with the activation solutions of Examples 2 and 3 above is enhanced, and the activation solution makes the binding force between the silica gel membrane and nucleic acid have good uniformity and stability.

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Abstract

The invention belongs to the technical field of nucleic acid extraction, in particular to a silicon membrane activation liquor and an application thereof. The invention aims at providing a method for carrying out processing on the silicon membrane; an activation technique can be adopted to increase the combining force of the silicon membrane and the nucleic acid; besides, the combining force thereof has excellent uniformity and stability. The invention improves the biological utilizing degree of the nucleic acid and better exerts the function for extracting the nucleic acid of the invention; besides, the silicon membrane is coated inside an activator by the method, thus isolating the contact with the outside environment; therefore, the character is more stable and the application range thereof is enlarged so as to meet different production demands.

Description

technical field [0001] The invention belongs to the technology in the field of nucleic acid extraction, and in particular relates to a silica gel membrane activation solution and its application. Background technique [0002] The research and application of molecular biology are inseparable from the extraction and purification of nucleic acid. The effect of nucleic acid extraction directly affects the follow-up work and research, but the extraction and purification of nucleic acid has always been a time-consuming and tedious process. Many scholars at home and abroad have been exploring various methods of nucleic acid extraction and purification. [0003] Traditional nucleic acid preparation methods often use toxic organic solvents such as phenol / chloroform for extraction. These techniques have a wide range of applications and are suitable for most molecular biology research. However, these techniques are cumbersome and time-consuming, and organic solvents such as phenol and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C07H1/06
Inventor 李晓晨
Owner TIANGEN BIOTECH BEIJING
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