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Genome-wide Alu element detection technique

A genome sequence and base technology, applied in the field of genome sequence detection, can solve the problems of cumbersome operation, inability to capture Alu subfamily, and many steps.

Inactive Publication Date: 2016-04-27
BGI SHENZHEN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Because this method uses AluYb8 / 9 specific primers to amplify, it can only capture about 1 / 3 of the Alu sequence, but cannot capture all the Alu subfamily
Moreover, the method has many steps, cumbersome operation, and takes a long time

Method used

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  • Genome-wide Alu element detection technique
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  • Genome-wide Alu element detection technique

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Embodiment Construction

[0024] Embodiments of the present invention are described in detail below. The embodiments described below are exemplary only for explaining the present invention and should not be construed as limiting the present invention. If no specific technique or condition is indicated in the examples, it shall be carried out according to the technique or condition described in the literature in this field or according to the product specification. The reagents or instruments used were not indicated by the manufacturer, and they were all commercially available conventional products.

[0025] Nested PCR (NestedPCR) uses a pair of outer primers to amplify the target sequence to increase the amount of template, and then uses a pair of inner primers to amplify to obtain specific PCR bands. Semi-nested PCR (Semi-Nested PCR) is a special type of nested PCR, which is used to amplify fragments smaller than ordinary PCR. Semi-nested PCR uses an outer primer instead of an inner primer. Among t...

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Abstract

The invention provides a novel method. A primer is designed according to common sequences of most subfamilies of Alu (except for AluYd3, Aluyd3a1, AluJo and AluJb, lower than 1% in content, with the coverage of remaining Alu subfamilies up to higher than 99% in addition to inactive subfamilies in a genome, and sequences of nearly all Alu subfamilies are specifically captured using semi-nested PCR (polymerase chain reaction).

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, the present invention relates to the detection of genomic sequences. Background technique [0002] Alu element (Aluelement) is a kind of repeated sequence scattered in the genome, which belongs to non-autonomous retrotransposon, and the size of each copy is about 300 base pairs. It is generally believed that the Alu element is formed by inserting a sequence of retrotransposase into the genome. There are about one million copies of Alu elements in each person's genome, and there is an Alu insertion every 3,000 bases on average. Abnormal Alu insertions in the genome and abnormal splicing and recombination of Alu can lead to cancer. Currently, 31 subfamilies of Alu are included in dbRIP and RepBase databases. The most subfamilies in the known Alu family include AluY, AluYa5, AluYa8, AluYb8 and AluYb9, among which AluYa5 accounts for more than 50% of Alu. According to the c...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 徐礼钦李贵波蒋润泽何家伟
Owner BGI SHENZHEN CO LTD