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Primer, probe, kit and detection method for SNP site polymorphism of ABCB1 gene C3435T

A technology of site polymorphism and kit, which is applied in the field of biomedical clinical molecular detection, can solve the problems of high cost, slow detection speed, and low detection accuracy of kits, achieve good specificity, reduce design difficulty, overcome less sensitive effects

Active Publication Date: 2022-02-25
郑州华沃生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, the present invention provides a primer, a probe, a kit and a detection method for the SNP site polymorphism of ABCB1 gene C3435T, to solve the slow detection speed caused by the method for determining the genotype using a relatively poor Ct method , the technical problems of low detection accuracy and high cost of the kit

Method used

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  • Primer, probe, kit and detection method for SNP site polymorphism of ABCB1 gene C3435T
  • Primer, probe, kit and detection method for SNP site polymorphism of ABCB1 gene C3435T
  • Primer, probe, kit and detection method for SNP site polymorphism of ABCB1 gene C3435T

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Embodiment 1

[0034]Example 1: A primer and probe for the SNP site polymorphism of ABCB1 gene C3435T, including molecular beacon sequences, ARMS primer sequences of wild-type molecular beacons, ARMS primer sequences of mutant molecular beacons, and universal downstream primers; The molecular beacon sequence includes a wild-type molecular beacon and a mutant molecular beacon; the nucleotide sequence of the wild-type molecular beacon is shown in SEQ.ID.NO.1; the nucleotide sequence of the mutant molecular beacon is shown in SEQ.ID.NO.1 .ID.NO.2 shown;

[0035] The ARMS primer sequence of the wild-type molecular beacon includes w1, w2, w3, w4 and w5; the nucleotide sequence of w1 is shown in SEQ.ID.NO.3; the nucleotide sequence of w2 is shown in SEQ.ID. Shown in NO.4; The nucleotide sequence of w3 is shown in SEQ.ID.NO.5; The nucleotide sequence of w4 is shown in SEQ.ID.NO.6; The nucleotide sequence of w5 is shown in SEQ.ID .NO.7 shown;

[0036] The ARMS primer sequence of the mutant molecul...

Embodiment 2

[0041] Embodiment 2: the difference with embodiment 1 is:

[0042] The ARMS primer sequence of the wild-type molecular beacon includes one of w1, w2, w3, w4 and w5; the ARMS primer sequence of the mutant molecular beacon includes one of m1, m2, m3, m4 and m5; the The nucleotide sequence of the universal downstream primer is shown in SEQ.ID.NO.13.

Embodiment 3

[0043] Embodiment 3: the difference with embodiment 1 is:

[0044] The ARMS primer sequence of the wild-type molecular beacon includes any two of w1, w2, w3, w4 and w5; the ARMS primer sequence of the mutant molecular beacon includes one of m1, m2, m3, m4 and m5 ; The nucleotide sequence of the universal downstream primer is shown in SEQ.ID.NO.13.

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Abstract

The invention provides a primer, a probe, a kit and a detection method for SNP site polymorphism of an ABCB1 gene C3435T, and belongs to the technical field of biomedical clinical molecular detection, in particular to the primer and the probe for the SNP site polymorphism of the ABCB1 gene C3435T. The primer comprises a molecular beacon sequence, an ARMS primer sequence of a wild type molecular beacon, an ARMS primer sequence of a mutant type molecular beacon and a universal downstream primer; the molecular beacon sequence comprises a wild type molecular beacon and a mutant type molecular beacon. According to the invention, SNP genotyping can be rapidly realized in one tube.

Description

technical field [0001] The invention relates to the technical field of biomedical clinical molecular detection, in particular to a primer, a probe, a kit and a detection method for the SNP site polymorphism of ABCB1 gene C3435T. Background technique [0002] Molecular beacon technology is a molecular probe with a neck-loop configuration based on the principle of nucleic acid base pairing and fluorescence resonance energy, first proposed by Tyagi and Kramer in 1996. During the annealing stage of PCR amplification, the molecular beacon and The generated target sequence binds and emits fluorescence. In the extension stage, it breaks away from the target sequence without interfering with the amplification. As the number of cycles increases, the amount of molecular beacons bound to the template also increases, and the final fluorescence intensity is proportional to the amount of amplified template. Proportional. The technology has extremely high specificity, and is easy to opera...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6886C12Q1/6858C12N15/11C12R1/92
CPCC12Q1/701C12Q1/6886C12Q1/6858C12Q2600/156C12Q2600/136C12Q2531/113C12Q2563/107
Inventor 王知丰秦付军吴书展
Owner 郑州华沃生物科技有限公司