A kit and method for detecting chromosome heterozygosity loss based on amplicon sequencing

A loss of heterozygosity, kit technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of chromosome deletion, non-disclosure and implication, high and low gene expression, etc. Achieve the effect of increasing the amount of DNA and reducing the cost of detection
CN109504770BActive Publication Date: 2019-11-19艾普拜生物科技(苏州)有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
艾普拜生物科技(苏州)有限公司
Publication Date
2019-11-19

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Abstract

The invention discloses a kit and method for detecting chromosome heterozygosity loss on the basis of amplicon sequencing, belongs to the field of molecular biology nucleic acid detection, and relatesto the kit and method for detecting chromosome heterozygosity loss by means of amplicon sequencing. The kit contains a mixture of 14 groups of primer pairs used for detecting a 1p36.32 region of a human chromosome 1, 6 groups of primer pairs used for detecting a 1q25.2 region of the human chromosome 1, 14 groups of primer pairs used for detecting a 19p13.2 region of a human chromosome 19 and 7 groups of primer pairs used for detecting a 19q13.33 region of the human chromosome 19, 20 kinds of tag primers used for distinguishing samples, a GC stabilizer, a PCR enzyme mixture, purifying magneticbeads, a quality control standard substance and ddH2O. Through two-time PCR amplification and two-time product purification and sequencing, through a ratio of READS(1p) / READS(1q) to READS(19q) / READS(19p), the heterozygosity loss state of 1p19q in the samples is analyzed. By adopting the method, the variation situation of the 1p19q can be reflected comprehensively, truly and accurately, and gene variation of the 96 samples can be subjected to high-throughput detection and analysis simultaneously, so that the detection cost is greatly reduced.
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Description

technical field

[0001] The invention belongs to the field of nucleic acid detection in molecular biology, and relates to a kit and a method for detecting chromosome heterozygosity loss based on an amplicon sequencing method. Background technique

[0002] Glioma originating from glial cells is the most common primary intracranial tumor, accounting for about 27% of all central nervous system tumors and about 80% of malignant tumors. The annual incidence of glioma in my country is (3-6.4) / 100,000, and the annual death toll reaches 30,000. The incidence rate of malignant glioma (Glioblastoma, GBM, WHO grade IV) is 5.8 / 100,000, and the 5-year mortality rate is second only to pancreatic cancer and lung cancer among systemic tumors, ranking third. Oligodendroglioma accounts for 5-18% of glioma, and is an independent type of glioma. Most oligodendroglial tumors have loss of heterozygosity (Loss of Heterozygosity, LOH) of the short arm of chromosome 1 (1p) and the long arm of chrom...

Claims

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