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HLA gene amplification primer, kit, sequencing library construction method and sequencing method

A technology for gene amplification and gene sequencing, applied in the field of HLA gene amplification primers, to achieve the effects of long read length accuracy, high accuracy, and simplified process flow

Pending Publication Date: 2021-12-21
XIAN HAORUI GENE TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Based on this, it is necessary to solve the problem of ambiguous results introduced by traditional HLA genotyping methods, and provide an HLA gene amplification primer, kit, sequencing library construction method and sequencing method that can improve the accuracy of the results and simplify the process.

Method used

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  • HLA gene amplification primer, kit, sequencing library construction method and sequencing method
  • HLA gene amplification primer, kit, sequencing library construction method and sequencing method
  • HLA gene amplification primer, kit, sequencing library construction method and sequencing method

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Embodiment 1

[0075] 1. Amplification

[0076] Amplification was performed using 88 human gDNA standard samples purchased from Coriell. The general adaptability and sensitivity of the method of the present invention are demonstrated by the results of a large number of samples.

[0077] Design specific amplification primers for different HLA genes, select 5’P for primer synthesis, and the following table 1 is the primer sequence:

[0078] Table 1

[0079]

[0080]

[0081] Primers are configured and diluted using EB buffer. First, dilute F and R primers respectively. After mixing F and R, configure primer MIX according to the primer ratio in Table 2 below.

[0082] Table 2

[0083]

[0084] After configuring the primer MIX, add the PCR amplification system according to the following table 3 in order, close the tube cap, mix well and centrifuge.

[0085] table 3

[0086]

[0087] Set the program of the PCR instrument according to the parameters in Table 4 below for amplificat...

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Abstract

The invention discloses an HLA (human leukocyte antigen) gene amplification primer. The HLA gene amplification primer comprises a mixture consisting of any one or more of the following nine primer groups of SEQ ID NO: 1 and SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6; SEQ ID NO: 7 and SEQ ID NO: 8; SEQ ID NO: 9 and SEQ ID NO: 10; SEQ ID NO: 11 and SEQ ID NO: 12; SEQ ID NO: 12 and SEQ ID NO: 13; SEQ ID NO: 14 and SEQ ID NO: 15; SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18. The invention also discloses a kit for sequencing the HLA gene, and the kit comprises the HLA gene amplification primer. The invention also discloses a construction method of a gene sequencing library based on the HLAHLA amplification primer. The invention also discloses an HLA gene sequencing method based on the construction method of the HLA gene sequencing library.

Description

technical field [0001] The invention relates to the technical field of nucleic acid sequencing, in particular to HLA gene amplification primers, a kit, a sequencing library construction method and a sequencing method. Background technique [0002] HLA (Human Leukocyte Antigen) molecules present antigens to T lymphocytes for recognition, and play an extremely important role in adaptive immune responses. HLA is a highly polymorphic complex composed of a series of closely linked gene loci. It is a gene cluster encoding human major histocompatibility complex (MHC). HLA is located on the short arm of chromosome 6. This system is the most complex polymorphic system known to the human body. HLA is a highly polymorphic alloantigen, and its chemical essence is a kind of glycoprotein, which is composed of an α heavy chain (glycosylated) and a β light chain non-covalently combined. The amino terminal of the peptide chain is outward (accounting for about 3 / 4 of the whole molecule), th...

Claims

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Application Information

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IPC IPC(8): C12N15/11C40B50/06C12Q1/6869
CPCC12N15/11C40B50/06C12Q1/6869C12Q2531/113C12Q2537/143C12Q2535/122C12Q2547/101
Inventor 王博林奇翰杨龙张悦
Owner XIAN HAORUI GENE TECH LTD
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