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Probe set and application thereof

A probe set and probe technology, applied in the biological field, can solve the problems of high price, long time-consuming, high detection cost, etc., and achieve the effect of family linkage detection

Pending Publication Date: 2019-12-03
MGI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the above-mentioned technology has the following disadvantages: PCR technology needs to screen the sites to be tested, design primers and adjust the experiment, which takes a long time in the early stage, and it is unavoidable during the detection process due to amplified allele dropout (ADO) The impact of the rate; SNP array technology has been initially used in clinical practice due to its fast, accurate and high-resolution characteristics, but the high price and analysis difficulties caused by excessive information in the detection process are the shortcomings of this technology
[0008] Therefore, establishing a liquid-phase capture probe combined with BGISEQ-500 PGD technical solution can overcome the shortcomings of the existing technology, such as long time-consuming and high detection costs in the early stage, so as to solve most of the genetic defects. the meaning of

Method used

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  • Probe set and application thereof
  • Probe set and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0191] The preparation of embodiment 1 probe set

[0192] This embodiment provides a set of probe sets, which are designed according to the following target genes, the target genes include: HBA1, HBA2, HBB, BCKDHA, CYP21A2, GFM1, ATP7B, GAA, IDS, DMD, ATXN1, ATXN2, ATXN7, CACNA1A, PKD1, PKD2, PKHD1, COL4A5, GUCY2D, USH2A, F8, F9, SH2D1A, PAH, AR, GALT, CFTR, SMN1, SMN2, HTT, CYBB, PRPF31, GJB2, GJB3, SLC26A4, NF1, NF2, BRCA1, BRCA2, RET, FBN1, FMR1, VHL, COL1A1, COL1A2, TSC1, MECP2 and HLA;

[0193] Each probe in the probe set was obtained according to the following probe design principles:

[0194] (1) The length of the probe is 90nt;

[0195] (2) When designing the probe, the window is slid at an interval of 1bp;

[0196] (3) removing probes containing unknown base sequences;

[0197] (4) When the ratio of multiple probes belonging to repetitive sequences is 100%, remove the multiple probes;

[0198] (5) Ensure that the probe coverage of each target area is 2X;

[0199...

Embodiment 2

[0215] The composition of embodiment 2 system

[0216] (1) Further, the inventor also provides a device for constructing a high-throughput sequencing library using the probe set in Example 1. The specific device structure is as follows: figure 2 As shown, the device includes a break repair unit, an adapter ligation unit, a first amplification unit, a hybridization elution unit, a second amplification unit and a circularization unit, specifically as follows:

[0217] A unit for obtaining genomic DNA, the unit for obtaining genomic DNA is connected to the interruption and repair unit for obtaining genomic DNA from a sample;

[0218] A break repair unit, the break repair unit is connected to the acquisition of genomic DNA unit, used for break repair of genomic DNA, to obtain DNA fragments;

[0219] A joint connection unit, the joint connection unit is connected with the interrupted repair unit, used for joint connection, and a label joint is added to obtain a connection product...

Embodiment 3

[0234] Example 3 Collection and analysis of α-thalassemia disease samples

[0235] 1) Sample collection

[0236] The family in this example is an α-thalassemia disease family, an autosomal recessive genetic disease, and the specific steps for collecting samples are as follows:

[0237] (1) The extratrophoblast cells of embryonic D3 blastomeres or D5 blastocysts are collected and subjected to multiple replacement expansion:

[0238] Collection: When the fertilized egg develops to the blastomere stage on the third day, one embryonic cell is biopsied through the zona pellucida, washed and placed in a PCR tube containing 4 μL of cell preservation solution; or when the fertilized egg develops to the fifth day At the blastocyst stage, a small piece of embryonic cells is biopsied by laser punching under a micromanipulator, washed and placed in a PCR tube containing 4 μL of cell preservation solution, and briefly centrifuged; the PCR tube with cells can be directly processed For who...

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Abstract

The invention relates to the biotechnology field, in particular to a probe set and application thereof. More particularly, the invention relates to one group of probe set, a method for constructing ahigh-throughput sequencing library, a method for determining single nucleotide mutation and haplotype of a sample to be detected, a device for constructing the high-throughput sequencing library and asystem for determining the single nucleotide mutation and the haplotype of the sample to be detected. The probe set can simultaneously detect the gene situations of various single gene inheritance diseases or genetic tumors, has good capture specificity, high sensitivity and wide coverage rate and can effectively realize the detection of 34 types of PGD (Preimplantation Genetic Diagnosis).

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, the present invention relates to a set of probe sets and their application. More specifically, the present invention relates to a set of probe sets, a method for constructing a high-throughput sequencing library, and determining the single-nucleus A method for nucleotide mutation and haplotype, a device for constructing a high-throughput sequencing library, and a system for determining the single nucleotide mutation and haplotype of a sample to be tested. Background technique [0002] Preimplantation Genetic Diagnosis (PGD) refers to the formation of embryos in the process of in vitro fertilization (IVF). Before the embryos are implanted in the uterus, embryos with known genetic risks of diseases or HLA matching requirements are tested. Biopsy and genetic analysis to select embryos without genetic diseases or in line with the expected human leukocyte antigen (HLA) type for impla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/6869C12N15/11C40B50/06
CPCC12Q1/6886C12Q1/6883C12Q1/6869C40B50/06C12Q2600/156C12Q2600/16C12Q2535/122
Inventor 谢林陈大洋刘萍殷旭阳夏军史千玉邱咏朱珠
Owner MGI TECH CO LTD
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