A method for simultaneous gene locus, chromosome and linkage analysis

Abstract

The invention relates to a method for simultaneously completing gene loci, chromosomes and linkage analysis, specifically including the acquisition of embryonic cell samples, whole genome amplification, target gene mutation site amplification, whole genome amplification products and target gene mutation sites The main steps of database construction, high-throughput sequencing and data analysis, through the use of whole genome amplification technology combined with high-throughput sequencing, multiple comprehensive tests can be completed in one step, avoiding the use of multiple methods and steps for single-gene genetic diseases Detection of mutation sites, chromosomal disorders and linkage analysis. The method provided by the present invention provides favorable conditions for micro-sample, not only can be used for PGD detection to determine whether the embryo carries a disease-causing gene and abnormal chromosome copy number; it is also suitable for genetic screening of embryos of recurrent miscarriages and elderly women, realizing a Steps to complete multiple detections of a single sample. Because of its simple operation, short period and strong feasibility, it is beneficial to popularization and application.

Description

technical field [0001] The invention relates to the fields of genome sequence analysis and bioinformatics, and specifically relates to a method for completing single-cell single-gene disease, chromosomal disease and linkage analysis in one step by using MALBAC amplification technology combined with high-throughput sequencing. Background technique [0002] Genetic diseases refer to diseases caused by changes in the genetic material (gene sequences on chromosomes or mitochondrial DNA) in the human body. In recent years, the incidence of genetic diseases has increased year by year. In my country, as many as tens of thousands of children with chromosomal abnormalities are born every year, and children with chromosomal abnormalities are still incurable. At present, there are more than 7,000 kinds of single-gene genetic diseases that have been discovered, and more than 4,000 of which have been identified as pathogenic genes. The overall morbidity rate in infants and the overall p...

AI Technical Summary

Problems solved by technology

Multiplex fluorescent PCR technology combines multiplex PCR with fluorescent probes to detect STR typing. Due to the lack of STR linkage markers, there may be no available STR sites in specific cases. Therefore, a lot of pre-experimental work is required before clinical testing. STR genetics Most of the markers are far away from the disease-causing site, prone to chromosomal recombination and misdiagnosis, and the detection cost is high, so this technology cannot be used for batch detection

Compared with the method of capturing SNP by chips and probes, the cost is relatively high and the cycle is relatively long compared with the method of capturing SNP by ordinary PCR

[0005] Existing technology cannot simultaneously complete the three items of monogenic genetic disease, chromosomal abnormality and linkage analysis through one-step detection, and the whole genome sample obtained from a small number of cells is prone to allelic dropout (ADO) when multiple tests are performed separately during the experiment , pollution and other phenomena, this field is in urgent need of a technical solution that can solve this problem

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