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Method for constructing variable region sequencing library, and method for determining variable region nucleic acid sequence

A technology for sequencing libraries and nucleic acid sequences, applied in the biological field, can solve problems such as MRD needs to be improved, and achieve the effect of improving efficiency

Inactive Publication Date: 2016-04-20
BGI SHENZHEN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the current methods for MRD detection still need to be improved

Method used

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  • Method for constructing variable region sequencing library, and method for determining variable region nucleic acid sequence
  • Method for constructing variable region sequencing library, and method for determining variable region nucleic acid sequence
  • Method for constructing variable region sequencing library, and method for determining variable region nucleic acid sequence

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Experimental program
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Embodiment 1

[0090] According to the general method described above, according to the following steps, construct the variable region sequencing library of samples of lymphocytic tumor patients at different stages before and after treatment, determine the variable region nucleic acid sequence, and perform MRD detection and prognosis evaluation for this patient, as follows :

[0091] (1) Experimental process:

[0092] 1) Sample preparation

[0093] For 4 acute lymphoblastic leukemia (T-ALL) patients (respectively named as patient 1, patient 2, patient 3, and patient 4), before treatment, 8 days after treatment, 15 days after treatment, and 33 days after treatment And 2-3 months after treatment, collect a test sample—bone marrow blood, and extract whole genome DNA or total RNA for later use. Wherein, DNA or RNA is extracted from any tissue containing lymphocytes (the detection sample can be any tissue containing lymphocytes, such as bone marrow, peripheral blood, solid tissue, and cultured ...

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Abstract

The present invention provides a method for constructing a variable region sequencing library, and a method for determining a variable region nucleic acid sequence. The method for constructing the variable region sequencing library comprises: adopting a sample containing nucleic acid encoding the variable region as a template, amplifying the nucleic acid encoding the variable region so as to obtain a first amplification product, and adding a sequencing linker to at least one of both terminals of the first amplification product so as to obtain the variable region sequencing library, wherein the primers used by the amplifying comprise: a first primer comprising at least a nucleic acid molecule specifically recognizing the framework encoding sequence of the V region, and a second primer comprising at least a nucleic acid molecule specifically recognizing the encoding sequence of the J region or C region. With the method of the present invention, the variable region sequencing library can be effectively constructed.

Description

technical field [0001] The present invention relates to the field of biotechnology, specifically, the present invention relates to a method for constructing a variable region sequencing library and a method for determining the variable region nucleic acid sequence. Background technique [0002] Lymphocytic neoplasms are malignant diseases that seriously endanger human health caused by malignant clonal proliferation of lymphocytes, including lymphocytic leukemia, myeloma and various lymphomas. With the improvement of diagnosis and treatment technology, the diagnosis and treatment of lymphocytic tumors has achieved a qualitative leap. Among them, the detection of minimal residues plays a key role in the stratification of lymphocytic tumors, evaluation of curative effect, estimation of patient prognosis, guidance of treatment plan formulation, and improvement of patient survival and cure. [0003] Minimal residual disease (MRD, Minimal residual disease) refers to the residual ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B50/06C12Q1/68
Inventor 武靖华刘晓王长希张伟曾晓静杜元平
Owner BGI SHENZHEN CO LTD
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