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Rapid library building method

A rapid and typing method technology, applied in the fields of genetic engineering and molecular biology, can solve the problems of low efficiency of library construction and complicated steps, and achieve the effect of improving the efficiency of library construction and simplifying experimental steps.

Active Publication Date: 2015-04-01
SHENGZHEN CHINA GENE TECH COMPANY
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Problems solved by technology

[0005] The purpose of the present invention is to provide a rapid library construction method, SNP typing method and sequencing method, aiming to solve the problems of complicated steps and low efficiency of library construction in the prior art

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no. 1 example

[0051] like figure 1 As shown, the present invention proposes the first embodiment, a method for rapidly building a database, comprising the following steps:

[0052] A. PCR amplifies the sample to be sequenced to obtain an amplified product; at least one of the PCR primers used for PCR amplification in the PCR amplification process contains a breakable site or an excisable sequence;

[0053] B. Adding the amplified product to a cleavage-ligation reaction system for cleavage and ligation to obtain a library molecule containing adapter 1; the cleavage-ligation reaction system includes: ligase, cleavage agent, adapter 1 and ligation buffer;

[0054]The cleaving agent is used to specifically cut the cleavable site or excisable sequence in the amplification product to form the first sticky end;

[0055] The first adapter is a nucleic acid molecule, which contains a second sticky end that is completely complementary to the first sticky end.

[0056] It should be noted:

[0057] ...

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Abstract

The invention relates to the fields of genetic engineering and molecular biology, and provides a rapid library building method, an SNP (single nucleotide polymorphism) genotyping method and a sequencing method. The rapid library building method includes the following steps: A. a sample to be sequenced goes through PCR (polymerase chain reaction) amplification, so as to obtain an amplified product; during the PCR amplification process, at least one PCR primer in a PCR primer pair for PCR amplification contains a breakable site or a cleavable sequence; B. the amplified product is added to a cleavage-ligation reaction system for cleavage and ligation, so as to obtain a library molecule containing a first linker; the cleavage-ligation reaction system comprises a ligase, a clastogen, the first linker and a ligation buffer solution; the clastogen is used for specific cleavage of the breakable site or the cleavable sequence in the amplified product, so as to form a first cohesive end; the first linker is a nucleic acid molecule, and contains a second cohesive end capable of completely complementary pairing with the first cohesive end. The invention simplifies experiment steps, and improves the library building efficiency.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, and more specifically, relates to a rapid library building method, a SNP typing method and a sequencing method. Background technique [0002] At the present stage, the sequence determination of the target region by sequencing technology often requires the construction of library molecules. The method of constructing library molecules generally includes the following steps: 1) Obtain the nucleic acid molecule containing the target region to be determined by PCR amplification; The two ends of the nucleic acid molecule containing the target region to be determined are respectively connected with different linker molecules to obtain a sequencing library. However, in step 2), in order to realize the directional connection between the two adapter molecules and the nucleic acid molecule containing the target region to be determined, it is necessary to sequentially carry out the ...

Claims

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Application Information

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IPC IPC(8): C40B50/06C12Q1/68
Inventor 盛司潼钟茂春
Owner SHENGZHEN CHINA GENE TECH COMPANY
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