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Oligonucleotide library classification and assessment method based on capillary zone electrophoresis

A technology of oligonucleotide library and zone electrophoresis, which is applied in the field of biological separation and analysis, can solve the problems of long time consumption, reduce the time and operation steps of the screening process, and heavy workload, and achieve the effect of simple data processing method

Inactive Publication Date: 2014-05-07
BEIJING INSTITUTE OF TECHNOLOGYGY
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Problems solved by technology

[0006] Aiming at the traditional CE-SELEX technology with heavy workload, long time consumption and repeated screening, the purpose of the present invention is to provide a method for grading and evaluating oligonucleotide libraries based on capillary zone electrophoresis. The library is fractionated, and the secondary library collected at different migration time periods by capillary zone electrophoresis interacts with the same target molecule to find the secondary library with the strongest binding ability to the target molecule, and then perform CE-SELEX technology , to reduce the time and steps of the screening process

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  • Oligonucleotide library classification and assessment method based on capillary zone electrophoresis
  • Oligonucleotide library classification and assessment method based on capillary zone electrophoresis
  • Oligonucleotide library classification and assessment method based on capillary zone electrophoresis

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Embodiment

[0039] A method for grading and evaluating an oligonucleotide library based on capillary zone electrophoresis, the steps are as follows:

[0040] step one,

[0041] (1) Dissolve the oligonucleotide library in the running buffer solution to make a stock solution 1 with a concentration of 100 μM of the oligonucleotide library, and store it at -20°C for later use; then dilute the stock solution 1 into a stock solution with a concentration of 20 μM stock solution 2;

[0042] Among them, the capillary used for capillary zone electrophoresis is a quartz fused capillary, which is covered with a silica gel layer. The inner diameter of the capillary is 75 μm, the total length is 50.2 cm, the side length of the detection window is 0.2 cm, and the distance from the window to the sample outlet is 10 cm; Rinse with water and running buffer solution for 3 minutes each; after washing, transfer the stock solution 2 to the capillary zone electrophoresis sample bottle, and use pressure injecti...

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Abstract

The invention discloses an oligonucleotide library classification and assessment method based on capillary zone electrophoresis, and belongs to the field of creature isolation analysis. The method comprises the following steps of: step one, carrying out capillary zone electrophoresis on an oligonucleotide library, and obtaining a secondary library within an oligonucleotide library electrophoresis time range according to transfer time slicing collection; and step two, respectively mixing each secondary library with a homogeneous target molecule, carrying out capillary zone electrophoresis, and comparing the strong or weak of each secondary library and target molecule interaction, thus obtaining the strongest secondary library of the target molecule combining capacity. The classification and assessment method can be used for realizing the fractionation of complicated constituent oligonucleotide library, and obtaining the strongest secondary library of the target molecule combining capacity; and the strongest secondary library of the combining capacity is utilized as a next CE (capillary electrophoresis)-SELEX(systematic evolution of ligands by exponential enrichment) technical screening library, the screening range is reduced, and the screening period of an adaptation body is shortened.

Description

technical field [0001] The invention relates to a method for grading and evaluating an oligonucleotide library based on capillary zone electrophoresis, belonging to the field of biological separation analysis. Background technique [0002] The oligonucleotide library is a class containing 10 13 -10 15 A mixture of DNA molecules with different bases, which may contain one or several DNA molecules that bind to the target molecule with high specificity and high affinity. Nucleic acid aptamer (Aptamer) refers to an oligonucleotide sequence ligand with high affinity and specificity obtained by screening from a random oligonucleotide library through exponential enrichment ligand system evolution (SELEX) technology. [0003] Nucleic acid aptamers have high affinity, strong specificity, easy preparation and modification, and wide distribution of target molecules. At present, SELEX technology screening methods for nucleic acid aptamers mainly include automated SELEX, subtractive S...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
Inventor 屈锋周晓玫赵新颖李倩
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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