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Method for capturing nucleic acid fragment

A technology for capturing nucleic acids and fragments, applied in the fields of genetic engineering and molecular biology, can solve problems such as poor uniformity and poor capture effect, achieve good uniformity, avoid poor capture effect, and reduce production costs

Active Publication Date: 2012-12-19
盛司潼
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for creation of highly specific binding molecules called biosensors with unique label numbers on their surface. These bindings are used to detect small amounts of substances like DNA (described below). They work well even when there may only one type of target material being detected at once due to imperfections such as uneven distribution across different samples containing many similar materials. By adding these specialized chains onto this biofilm they allow for more precise detection than traditional methods without sacrificially increasing costs associated with each individual analysis process.

Problems solved by technology

This patented technical problem addressed in this patents relates to finding ways to effectively isolate small amounts or even single cells containing genetic material from complex mixtures such as blood samples without damaging them due to nonuniform distribution of capture agents at different locations within each sample.

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  • Method for capturing nucleic acid fragment
  • Method for capturing nucleic acid fragment
  • Method for capturing nucleic acid fragment

Examples

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no. 1 example

[0061] The present invention proposes a first embodiment, the nucleic acid molecule containing the nucleic acid fragment to be captured is a cloning carrier library, and there are multiple nucleic acid fragments to be captured and constructed into multiple cloning vectors respectively, and the step A includes the following steps:

[0062] A01. Use restriction enzymes to cut off the nucleic acid fragments to be captured in the cloning vector library to obtain DNA templates;

[0063] A02. Fragmenting the DNA template to obtain DNA template fragmentation products;

[0064] A03. The DNA template fragmentation product is connected with a biotin-labeled adapter element to obtain a biotin-labeled capture probe.

[0065] In this protocol, the nucleic acid fragment to be captured is digested from the cloning vector by enzyme digestion, and then connected with a biotin-labeled adapter element to obtain a biotin-labeled capture probe. The cloning vectors in the cloning vector library in...

no. 3 example

[0078] The present invention also proposes a third embodiment, the step A includes the following steps:

[0079] A21. Fragmentation of nucleic acid molecules containing nucleic acid fragments to be captured to obtain fragmentation products;

[0080] A22. The fragmentation product is connected with the adapter element to obtain a nucleic acid fragment containing the adapter;

[0081] A23. Perform PCR amplification on the product of step A22 using biotin-labeled amplification primers to obtain biotin-labeled capture probes.

[0082] It should be noted that the nucleic acid molecule containing the nucleic acid fragment to be captured is a PCR amplification product, and the PCR amplification product uses corresponding PCR primers to amplify the target region on genomic DNA, mitochondrial DNA, plasmid or RNA acquired. The target area is composed of nucleic acid fragments to be captured. There may be multiple target areas. The linker element is a nucleic acid molecule, used for ...

no. 4 example

[0087] The present invention also proposes a fourth embodiment, the step A includes the following steps:

[0088] A31. Fragmentation of nucleic acid molecules containing nucleic acid fragments to be captured to obtain fragmentation products;

[0089]A32. The fragmented product is connected to a biotin-labeled adapter element to obtain a biotin-labeled capture probe.

[0090] It should be noted that the nucleic acid molecule containing the nucleic acid fragment to be captured is a PCR amplification product, and the PCR amplification product uses corresponding PCR primers to amplify the target region on genomic DNA, mitochondrial DNA, plasmid or RNA acquired. The target area is composed of nucleic acid fragments to be captured. There may be multiple target areas. The ligation of the fragmented product with the biotin-labeled adapter element can only occur at one or both ends of the fragmented product of the DNA template.

[0091] The biotin-labeled linker element is a nuclei...

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Abstract

The invention relates to the fields of gene engineering and molecular biology and provides a method for capturing a nucleic acid fragment. The method comprises the following steps: A, preparing a capture probe by using biotin labeled amplimers or joint elements with nucleic acid molecules containing a to-be-captured nucleic acid fragment as a raw material; and B, hybridizing the capture probe with a to-be-captured nucleic acid fragment library and capturing the to-be-captured nucleic acid fragment by using a solid phase carrier containing a streptavidin or avidin mark. The method for capturing the nucleic acid fragment in the invention can avoid occurrence of poor capture effects caused by poor homogeneity of the capture probe and enables cost for capturing of nucleic acid fragments to be reduced.

Description

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Claims

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Application Information

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Owner 盛司潼
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