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apoB (apolipoprotein B) gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip

A detection solution and specific technology, applied in the field of molecular biology, can solve the problems of easy contamination of samples, low sensitivity and high false positive rate, and achieve consistent detection results, simple steps and good detection specificity.

Active Publication Date: 2012-07-04
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are all based on PCR technology, which has the disadvantages of low sensitivity, easy sample contamination, and high false positive rate. At the same time, due to the limitation of detection throughput, it cannot meet the needs of practical applications.

Method used

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  • apoB (apolipoprotein B) gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip
  • apoB (apolipoprotein B) gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip
  • apoB (apolipoprotein B) gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip

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Experimental program
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Embodiment 1

[0024] The apo B gene SNP detection liquid chip mainly includes:

[0025] 1. ASPE Primers

[0026] Specific primer sequences were designed for wild-type and mutant types of five common genotypes of apo B gene, C181T, C245A / T, G153A, G82A and G62A. ASPE primers consist of "Tag sequence + specific primer sequence". ASPE primer sequences are shown in the table below:

[0027] Table 1 ASPE primer sequence of apo B gene (Tag sequence + specific primer sequence)

[0028]

[0029]

[0030] Each ASPE primer includes two parts, the 5' end is a specific tag sequence for the anti-tag sequence on the corresponding microsphere, and the 3' end is a mutant or wild-type specific primer fragment (as shown in the above table 1). All ASPE primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Each synthesized primer was prepared into a 100pmol / mL stock solution with 10mmol / LTris Buffer.

[0031] 2. Microspheres coated with anti-tag sequences

[0032] ...

Embodiment 3

[0112] The detection of the liquid phase chip of embodiment 3 different ASPE primers to apo B gene SNP site

[0113] 1. Design of liquid phase chip preparation (selection of tag sequence and anti-Tag sequence)

[0114] Taking the liquid phase chip for detection of site mutations of apo B gene C181T and G153A as an example, the specific primer sequences at the 3' end of the ASPE primers were designed for the wild type and mutant types of C181T and G153A, respectively, and the tag sequences at the 5' end of the ASPE primers were selected from SEQ ID NO.1-SEQ ID NO.11, correspondingly, the anti-tag sequence coated on the microsphere and complementary to the corresponding tag sequence is selected from SEQ ID NO.23-SEQ ID NO.33. The specific design is shown in the following table (Table 7). The synthesis of ASPE primers, microspheres coated with anti-tag sequences, amplification primers, detection methods, etc. are as described in Example 1 and Example 2.

[0115] Table 7 Design ...

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Abstract

The invention discloses an apoB gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip. The liquid-phase chip comprises an ASPE (Allele Specific Primer Extension) primer, microspheres and an amplification primer, wherein the ASPE primer consists of a tag sequence at 5' end and specific primers aiming at target gene mutation at 3' end, wherein the specific primers are SEQ ID NO. 12 and SEQ ID NO. 13 aiming at a C181T SNP site, SEQ ID NO. 14 and SEQ ID NO. 15 aiming at a C245A SNP site, SEQ ID NO. 14 and SEQ ID NO. 15 aiming at a C245T SNP site, SEQ ID NO. 17 and SEQ ID NO. 18 aiming at a G153A SNP site, SEQ ID NO. 19 and SEQ ID NO. 20 aiming at a G82A SNP site, and / or SEQ ID NO. 21 and SEQ ID NO. 22 aiming at a G62A SNP site; and the microspheres are coated by anti-tag sequence. The matching ratio of the detection result of the apoB gene SNP detection liquid-phase chip provided by the invention and that of a sequencing method reaches up to 100 percent; and parallel detection of a plurality of polymorphic sites is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for detecting apo B gene SNP and a liquid phase chip. Background technique [0002] Apolipoprotein B (apolipoprotein B, apoB) is an important component of lipoproteins rich in cholesterol and triglyceride, and plays an important role in the transport, metabolism and maintenance of lipid levels in the body. Apolipoprotein B has two isoforms: apo B 100 and apo B 48. Apo B 48 is produced in the small intestine and is only found in CM. It is the N-terminal part of apo B100; The assembly, secretion, and degradation of lipoproteins containing apo b are closely related, and play an important role in maintaining a constant blood lipid level in the body, and its increase is one of the main risk factors for atherosclerosis. [0003] The apo B gene is located on the short arm of human chromosome 2, with a total length of 43kb, conta...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 许嘉森秦会娟郭婧朱泽尧
Owner SUREXAM BIO TECH
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