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Primer pair, probe and kit for detecting SLCO1B1 521T)C gene polymorphism

A technology of SLCO1B1521T and SLCO1B1521, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of long detection time and obvious non-specific amplification, so as to facilitate clinical use and reduce non-specific amplification. The effect of heterosexual amplification and shortening detection time

Inactive Publication Date: 2018-12-11
东莞博盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a pair of primers, probes and kits for detecting SLCO1B1 521T>C gene polymorphism, so as to solve the problem of long time-consuming detection of SLCO1B1 521T>C gene polymorphism and non-specific amplification in the prior art. problems such as increased

Method used

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  • Primer pair, probe and kit for detecting SLCO1B1 521T)C gene polymorphism
  • Primer pair, probe and kit for detecting SLCO1B1 521T)C gene polymorphism
  • Primer pair, probe and kit for detecting SLCO1B1 521T)C gene polymorphism

Examples

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Embodiment 1

[0032] Example 1. Primer pair for detecting SLCO1B1 521T>C gene polymorphism and preparation method thereof

[0033] In this example, a primer pair was designed for the 521 site of SLCO1B1.

[0034] According to rs4149056 sequence (SEQ ID No.5) design primer pair, primer pair sequence is as follows:

[0035] Forward primer (SEQ ID No.1): 5' (Amino group)- GGTCATACATGTGGAT3';

[0036] Reverse primer (SEQ ID No.2): 5' (Amino group)- TCTCCCCTATTCCAC 3'.

[0037] In this embodiment, the preparation of primers includes: designing primers with base sequences such as SEQ ID No.1 and SEQ ID No.2 for the 521 site of SLCO1B1, and using an amino group to base the 5' end of the base sequence base modification, that is.

[0038] In the process of preparing the above primer pairs, the Tm values ​​of the base sequence before the amino group modification and the base sequence after the amino group modification were tested by using the primer design software primer express of ABI comp...

Embodiment 2

[0043] Example 2. Probe for detecting SLCO1B1 521T>C gene polymorphism and its preparation method

[0044] In this example, a fluorescent probe was designed for the 521 site of SLCO1B1.

[0045] Design wild type probe and mutant probe according to rs4149056 sequence (SEQ ID No.5) mutation fragment:

[0046] Wild type probe (SEQ ID No.3):

[0047] 5'FAM-TAT (amino group) G- T -GTTCATGGGTAATATGC-BHQ1 3', the third base (T) from the 5' end is modified with an amino group, and the fifth base T from the 5' end is the SNP site, which is wild type.

[0048] Mutant probe (SEQ ID No.4):

[0049] 5'Cy5-FAM-TATG- C -G (amino group) TTCATGGGGTAATATGC-BHQ2 3';

[0050] The sixth base (G) from the 5' end is modified with an amino group, and the fifth base C from the 5' end is the SNP site, which is a variant.

[0051] The preparation of the above-mentioned fluorescent probe includes: designing a base sequence for the 521 site of SLCO1B1 as a fluorescent probe shown in SEQ ID No.3 an...

Embodiment 3

[0056] Example 3. Kit for detecting SLCO1B1 521T>C gene polymorphism

[0057] In this embodiment, a kit containing the primer pair of Example 1 and the probe of Example 2 is designed for the 521 site of SLCO1B1.

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Abstract

The invention discloses a primer pair, a probe and a kit for detecting SLCO1B1 521T)C gene polymorphism. The kit comprises a primer and a fluorescence probe; the primer is designed for SLCO1B1 521T locus and has a base sequence shown as SEQ ID No.1 and SEQ ID No.2, wherein a 5'-base of the base sequence is equipped with an amino-modified group; the fluorescence probe is designed for SLCO1B1 521T locus and has the base sequence shown as SEQ ID No.3 and SEQ ID No.4, wherein one basic group in the base sequence is equipped with the amino-modified group. Compared with the prior art, the kit disclosed by the invention adopts the amino modified primer and probe, further limits the detection time and is capable of obviously shortening the time required by detection, reducing non-specific amplification, detecting SNPs and point mutation, reducing design difficulty and detection cost without influence on diagnosis accuracy, realizing a method for detecting mutation without opening a single reaction tube, extremely avoiding aerosol pollution, bringing convenience to clinic application and reducing the error caused by operation.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a pair of primers, a probe and a kit for detecting SLCO1B1 521T>C gene polymorphism. Background technique [0002] Polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. Fluorescent PCR (qPCR) detection is a real-time detection technology developed on the basis of PCR technology. It uses the accumulation of fluorescent signals to monitor the entire PCR process in real time. Finally, the standard curve is used to quantitatively analyze the unknown template or detect the target gene based on the accumulation of fluorescent signals. This technology has achieved a leap from qualitative to quantitative PCR, and compared with conventional PCR, it has the characteristics of stronger specificity, effective solution to the problem of PCR pollution, and high degree of automation. It has been widely used by the beginning of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/106C12Q2600/156C12Q2563/107
Inventor 金京勋刘昕王弋刘芳崔红
Owner 东莞博盛生物科技有限公司
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