Method for detecting gene mutation

A detection method and forward primer technology, which are applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of lack of gene mutation methods, poor detection, and difficult amplification, etc. The effect of detection cost, easy judgment, and short time

Active Publication Date: 2011-04-06
广东智威农业科技股份有限公司 +1
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AI Technical Summary

Problems solved by technology

At present, there are many literatures and patents reporting the detection method of some mutation sites on the gene, one of the important mutation sites (type I mutation) is a large fragment of 1781 bp located in exon 10 and 3' untranslated region Deletion, because the target band amplified by the deletion mutation site is relatively large (greater than 2000 bp), it is not easy to amplify, and the amplification is unstable, so it cannot be detected well
[0007]Currently there is a lack of methods that can detect both long deletion mutations and gene mutations in long deletion sequences

Method used

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  • Method for detecting gene mutation
  • Method for detecting gene mutation
  • Method for detecting gene mutation

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Embodiment Construction

[0026] Below in conjunction with embodiment, further illustrate the present invention.

[0027] chicken GHR Establishment of a new method for molecular detection of type I mutations in genes

[0028] (1) Primers:

[0029] take chicken GHR Design primers P1-F / R for the sequence at both ends of the type I mutation deletion fragment on the gene, and design primer P2-F / R according to the sequence at both ends of the type II mutation (reference sequence GeneBank accession number: AC187590), and amplify the obtained sequence and related information such as figure 1 shown. The primer sequences used are as follows:

[0030] P1-F: 5'-GGCAGAAACCCCAAGTATG-3' (SEQ ID NO: 1);

[0031] P1-R: 5'-TCGGGACAGATCAAAGACAAT-3' (SEQ ID NO: 2);

[0032] P2-F: 5'-ATCCTGTTGCTTTCTTATGG-3' (SEQ ID NO: 3);

[0033] P2-R: 5'-CTCAGTCCTGTAGCCCTTG-3' (SEQ ID NO: 4).

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Abstract

The invention discloses a method for detecting gene mutation. The method comprises the following steps of: synthesizing a forward primer P1-F and a reverse primer P1-R of a long segment deficiency gene mutation sequence; synthesizing a forward primer P2-F and a reverse primer P2-R of a gene mutation sequence of the long segment deficiency gene mutation sequence; using a sequence to be detected as a template and performing a polymerase chain reaction (PCR) for one time by using a mixture of the four primers as a primer; and performing genotype analysis on the product of the PCR to determine the condition of the gene mutation. By the method of the invention, the long segment sequence deficiency mutation and the gene mutation thereof can be detected conveniently at the same time. By the method for detecting the gene mutation, the long segment sequence deficiency mutation can be detected more conveniently and stably without using expensive reagents such as linker adaptor (LA) amplification enzyme and the like; the cost of detecting the gene mutation is reduced; and the detection efficiency is improved.

Description

technical field [0001] The invention relates to a detection method for gene mutation, in particular to a detection method capable of simultaneously detecting two mutant genes. Background technique [0002] Known gene mutations include long-segment deletion mutations, and other gene mutations may also exist in long-segment deletion mutations. The confirmation of these gene mutations, especially in the case of the same external phenotype of these gene mutations, has important practical significance. [0003] Long fragment deletions are generally detected by long PCR method, which is limited to the existing PCR technology. The efficiency of long chain PCR is low, the amplification is unstable, and it is prone to errors. Therefore, this detection method has certain limitations. Due to the need to use expensive The detection cost of special polymerases also remains high. In addition, in order to detect different gene mutations at the same time, it generally needs to be carried ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 罗成龙王艳邓科源陈顺艳彭志李春雨杨纯芬舒鼎铭
Owner 广东智威农业科技股份有限公司
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