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Multiplex PCR primer for amplifying human EGFR gene and design method thereof

A design method and primer design technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of complex amplification system, fragment loss, low amplification efficiency, etc., and achieve improved amplification Increase efficiency, simplify procedures, and reduce costs

Active Publication Date: 2013-03-06
陕西佰美医学检验有限公司
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Problems solved by technology

[0014] In order to solve the technical problems of fragment loss, low amplification efficiency and complicated amplification system in the design method of primers for multiplex PCR for amplifying human EGFR gene, the invention provides a multiplex PCR primer for amplifying human EGFR gene and its design method

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  • Multiplex PCR primer for amplifying human EGFR gene and design method thereof
  • Multiplex PCR primer for amplifying human EGFR gene and design method thereof
  • Multiplex PCR primer for amplifying human EGFR gene and design method thereof

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Embodiment Construction

[0058] A set of primers used in multiplex PCR should bind specifically to the target sequence and should not interfere with each other so as to amplify a sufficient amount of the target sequence. Multiplex PCR can amplify multiple target genes at the same time, which has the advantages of saving time, reducing costs, and improving efficiency, especially saving precious experimental samples. Since multiplex PCR requires the specific amplification of multiple sites in the same reaction system, the technical difficulty increases. An ideal multiplex PCR reaction system is not a simple mixture of single PCRs. It requires comprehensive analysis and repeated experiments for the target product to establish a suitable reaction system and reaction conditions. A large number of experiments have shown that the main technical problem of multiplex PCR lies in the design, combination and concentration optimization of primers. The current research is mainly carried out in two aspects in the ...

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Abstract

The invention relates to a multiplex polymerase chain reaction (PCR) primer for amplifying a human epidermal growth factor receptor (EGFR) gene and a design method thereof. The design method comprises the following steps of: 1, designing a primer library of the human EGFR genes by using primer design software; 2, screening the primers in the primer library; 3, judging advantage and disadvantage of competitiveness of retained primers, performing the step 4 for the primers with competitiveness disadvantage, and performing the step 5 for the primers with competitiveness advantage; step 4, rescreening the primers with competitiveness disadvantage; 5, amplifying the primers with competitiveness advantage by using a term-PCR method; and 6, removing redundant primers in amplification primers to obtain the multiplex PCR primer. In the invention, the technical problems of segment loss, low amplification efficiency and complex amplification system existing in the design method of the conventional primer are solved; and the primer and the design method have the advantages of reducing the phenomenon of segment loss in the multiplex PCR system and optimally utilizing the resource.

Description

technical field [0001] The invention relates to the field of multiplex PCR of human epidermal growth factor EGFR (Epidermal growth factor receptor, EGFR) gene, in particular to a primer for multiplex PCR of human EGFR gene and a method thereof. Background technique [0002] Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors in the world today. Despite the continuous improvement of surgery and chemotherapy techniques, the prognosis of patients with advanced lung cancer is still poor. In recent years, with the improvement of molecular technology, anti-tumor drugs targeting molecular targets have been produced. These drugs are highly targeted, can specifically kill tumor cells, and have remarkable effects. Among them, the molecular targeted therapy targeting epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors is becoming more and more prominent in the treatment of non-small cell lung cancer. This kind of targeted drug is called tyrosin...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 陈超饶文波姬婧朱娟莉宗廷勇李颖朱雅清訾静张健杨莎莎王惠惠张锦超边婷张开元
Owner 陕西佰美医学检验有限公司
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